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1.
Article | IMSEAR | ID: sea-203729

ABSTRACT

Catharanthus roseus (periwinkles) belongs to the Apocynaceae family with great anti-cancer, anti-diabetic, andhepatoprotective values. Due to the large number of active molecules accumulated in these plants, they are ofparticular concern, especially in the pharmaceutical sector. The availability of ESTs gave the genetic algorithmof the plant to differentiate between the species accessions at the genetic level. The high-throughput method usedfor mining and detection of microsatellites (SSRs) embedded in ESTs gave a new insight for molecular markers’development. 19899 ESTs were retrieved, examined by NCBI EST dB and assembled 2692 to get full-lengthcontigs sequences. 338 microsatellites (SSR) loci were predicted with an average of SSR per 9.33 kb of ESTthough MISA-web tools out of 2692 contigs. Furthermore, trinucleotide, a well-known SSR was examined andfound to be the most favorable repeats' type (26.62%) followed by dinucleotide (24.22), mononucleotide (48.22%),and hexanucleotide (0.3%) types. The highest frequency of (A/T)n was reported in this finding followed by(AAG)n. The simple sequence repeats (SSR) extracted from C. roseus EST's data were used as molecular toolsfor genetic characterization in the present study. These predicted SSRs can be significantly used for constructingthe genetic maps and also for differentiating the accession between the species.

2.
Electron. j. biotechnol ; 39: 30-41, may. 2019. tab, ilus
Article in English | LILACS | ID: biblio-1051590

ABSTRACT

BACKGROUND: A total of 62,591 cowpea expressed sequence tags (ESTs) were BLAST aligned to the whole-genome sequence of barrel medic (Medicago truncatula) to develop conserved intron scanning primers (CISPs). The efficacy of the primers was tested across 10 different legumes and on different varieties of cowpea, chickpea, and pigeon pea. Genetic diversity was assessed using the same primers on different cowpea genotypes. Singlenucleotide polymorphisms (SNPs) were detected, which were later converted to length polymorphism markers for easy genotyping. CISPs developed in this study were used in tagging resistance to bacterial leaf blight disease in cowpea. RESULTS: A total of 1262 CISPs were designed. The single-copy amplification success rates using these primers on 10 different legumes and on different varieties of cowpea, chickpea, and pigeon pea were approximately 60% in most of the legumes except soybean (47%) and peanut (37%). Genetic diversity analysis of 35 cowpea genotypes using 179 CISPs revealed 123 polymorphic markers with PIC values ranging from 0.05 to 0.59. Potential SNPs identified in cowpea, chickpea, and pigeon pea were converted to PCR primers of various sizes for easy genotyping. Using the markers developed in this study, a genetic linkage map was constructed with 11 linkage groups in cowpea. QTL mapping with 194 F3 progeny families derived from the cross C-152 × V-16 resulted in the identification of three QTLs for resistance to bacterial leaf blight disease. Conclusions: CISPs were proved to be efficient markers to identify various other marker classes like SNPs through comparative genomic studies in lesser studied crops and to aid in systematic sampling of the entire genome for well-distributed markers at low cost


Subject(s)
Genome, Plant , Genomics/methods , Medicago truncatula/genetics , Polymerase Chain Reaction , Chromosome Mapping , Expressed Sequence Tags , Polymorphism, Single Nucleotide , Genomics , Quantitative Trait Loci , Fabaceae/genetics
3.
Indian J Exp Biol ; 2013 Jul; 51(7): 522-530
Article in English | IMSEAR | ID: sea-147623

ABSTRACT

Chinese cabbage (Brassica rapa) is widely recognized for its economic importance and contribution to human nutrition but abiotic and biotic stresses are main obstacle for its quality, nutritional status and production. In this study, 3,429 Express Sequence Tag (EST) sequences were generated from B. rapa cv. Osome cDNA library and the unique transcripts were classified functionally using a gene ontology (GO) hierarchy, Kyoto encyclopedia of genes and genomes (KEGG). KEGG orthology and the structural domain data were obtained from the biological database for stress related genes (SRG). EST datasets provided a wide outlook of functional characterization of B. rapa cv. Osome. In silico analysis revealed % 83 of ESTs to be well annotated towards reeds one dimensional concept. Clustering of ESTs returned 333 contigs and 2,446 singlets, giving a total of 3,284 putative unigene sequences. This dataset contained 1,017 EST sequences functionally annotated to stress responses and from which expression of randomly selected SRGs were analyzed against cold, salt, drought, ABA, water and PEG stresses. Most of the SRGs showed differentially expression against these stresses. Thus, the EST dataset is very important for discovering the potential genes related to stress resistance in chinese cabbage, and can be of useful resources for genetic engineering of Brassica sp.


Subject(s)
Brassica/drug effects , Brassica/genetics , Brassica/growth & development , Databases, Genetic , Expressed Sequence Tags/metabolism , Gene Expression Profiling , Gene Library , Gene Regulatory Networks , Genes, Plant/genetics , Genome, Plant , Humans , Molecular Sequence Annotation , Oligonucleotide Array Sequence Analysis , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction , Sodium Chloride/pharmacology , Stress, Physiological/genetics
4.
Rev. colomb. biotecnol ; 14(1): 191-199, ene.-jun. 2012. ilus, graf, tab
Article in Spanish | LILACS | ID: lil-656951

ABSTRACT

La identificación de genes nuevos relacionados con la respuesta de Nicotiana tabacum L. al estrés biótico y abiótico contribuye al mejoramiento genético del cultivo del tabaco en todo el mundo. El objetivo de este trabajo fue detectar la presencia de los genes tpt1 y pr1 en el genoma de algunas especies y variedades cubanas de tabaco. Se identificaron 265 etiquetas de secuencias expresadas (ESTs-expressed sequences tags) que nunca se habían informado con anterioridad en especies vegetales, y el gen que codifica para la proteí­na tumoral controlada durante la transcripción (Transcriptional Controlled Tumor Protein) (tpt1) nunca se había informado en N. tabacum, pues los estudios del mismo se han centrado en su mayoría en animales y humanos. Los resultados del microarreglo se confirmaron utilizando la RT-PCR cuantitativa en tiempo real. Los genes tpt1 y proteí­na relacionada con la patogénesis (pr1) se utilizaron para diseñar cebadores para la caracterización molecular de algunas especies y variedades de tabaco cubano. El gen tpt1 solamente se expresó en dos especies (N. glutinosa y N. tomentosiformis) y el pr1, después de la digestión, mostró diferentes bandas entre las variedades susceptibles y resistentes. La presencia de estos genes en una parte del germoplasma analizado constituye un paso inicial para la utilización de este conocimiento en el mejoramiento genético del tabaco.


The identification of new genes related with Nicotiana tabacum L. response to biotic and abiotic stress contribute to the genetic improvement of tobacco plants around the world. The aim of this research was to identify tpt1 and pr1 genes in the genomic of some cuban tobacco varieties and species. The microarray technology has been used with ESTs for the construction of a cDNA library. 265 expressed sequences tags (ESTs) that have never been reported before in vegetables species were identified and the gene that codified for the Transcriptional Controlled Tumor Protein (tpt1), has never been reported before in N. tabacum, with the studies about this gene being focused on human and animals mostly. The microarray results were confirmed using quantitative RT- PCR. TCTP and pathogenesis-related protein 1 (PR-1) ESTs were used to design primers for the molecular characterization of some species and cuban tobacco varieties. The tpt1 gene was only expressed in two species (N. glutinosa y N. tomentosiformis) and pr1 gen, after a digestion, exhibited different bands for susceptible and resistant varieties. The detection of these genes in some species and cuban tobacco varieties represents one step to go deeply in the molecular characterization of cuban germoplasm.


Subject(s)
Genes , Models, Molecular , Tabacum , Nicotiana , Molecular Structure
5.
J Biosci ; 2011 Jun; 36(2): 223-228
Article in English | IMSEAR | ID: sea-161536

ABSTRACT

Late embryogenesis abundant (LEA) protein family is a large protein family that includes proteins accumulated at late stages of seed development or in vegetative tissues in response to drought, salinity, cold stress and exogenous application of abscisic acid. In order to isolate peanut genes, an expressed sequence tag (EST) sequencing project was carried out using a peanut seed cDNA library. From 6258 ESTs, 19 LEA-encoding genes were identified and could be classified into eight distinct groups. Expression of these genes in seeds at different developmental stages and in various peanut tissues was analysed by semi-quantitative RT-PCR. The results showed that expression levels of LEA genes were generally high in seeds. Some LEA protein genes were expressed at a high level in non-seed tissues such as root, stem, leaf, flower and gynophore. These results provided valuable information for the functional and regulatory studies on peanut LEA genes.

6.
Rev. colomb. psicol ; 20(1): 99-106, jun. 2011.
Article in Spanish | LILACS | ID: lil-606142

ABSTRACT

En los últimos años han aumentado las investigaciones clínicas que utilizan el método científico para su validación, en gran parte influidas por el movimiento de los tratamientos con apoyo empírico (TAE) iniciado en la década de 1990. Se presentan los antecedentes de los TAE, sus objetivos y resultados, así como dos modelos de intervención que complementan a los TAE: las investigaciones sobre qué tipo de relación terapéutica predice más el éxito en las terapias, y las que buscan optimizar los sistemas de atención de salud mental. Se concluye que la mayoría de los TAE son técnicas que se basaron en la psicología experimental y que el método científico es la mejor herramienta con la que se cuenta para enfrentar el complejo problema de la conducta humana.


The last few years have seen an increase in clinical research that uses the scientific method for its validation, mainly due to the influence of the empirically supported treatments (ESTs) movement that started in the 1990s. The article describes the background, objectives, and results of ESTs, as well as two intervention models that complement them: one that promotes research on the type of therapeutic relation that guarantees greater success of the therapies, and one that seeks to optimize mental health care systems. The conclusion is that most ESTs are techniques based on experimental psychology, and that the scientific method is the best tool available to address the complex problem of human behavior.


Subject(s)
Psychology, Clinical , Psychotherapy , Psychology/methods , Psychology/trends , Scientific Research and Technological Development , Behavioral Medicine , Psychology, Applied
7.
Acta biol. colomb ; 16(1): 95-108, abr. 2011.
Article in Spanish | LILACS | ID: lil-635067

ABSTRACT

La yuca (Manihot esculenta) constituye la base de la alimentación para más de 1.000 millones de personas en el mundo, consolidándose como el cuarto cultivo más importante en el mundo después del arroz, el maíz y el trigo. La yuca es considerada como un cultivo relativamente tolerante a condiciones de estrés abiótico y biótico; sin embargo estas características se encuentran principalmente en variedades no comerciales. Las estrategias de mejoramiento genético convencional o mediadas por transformación genética representan una alternativa para introducir las características deseadas dentro de las variedades comerciales. Un paso fundamental con miras a acelerar los procesos de mejoramiento genético en yuca requiere el descubrimiento de los respectivos genes relacionados con las características buscadas, para lo cual los ESTs (del inglés Expressed Sequence Tags) son una vía rápida para este fin. En este estudio se realizó un análisis de la colección completa de ESTs disponibles en yuca, representada por 80.459 secuencias, los cuales fueron ensamblados en un conjunto de 29.231 genes únicos (unigen), representado por 10.945 contigs y 18.286 singletones. Estos 29.231 genes únicos pueden representar cerca del 80% de los genes del genoma de yuca. Entre el 5 y 10% de los unigenes de yuca no presentaron similitud con las secuencias presentes en las bases de datos de NCBI y pueden constituir genes específicos de yuca. A un grupo de secuencias del set unigen (29%) fue posible asignarles una categoría funcional de acuerdo al vocabulario Gene Ontology. El componente función molecular es el mejor representado con 43% de las secuencias, seguido por el componente proceso biológico (38%) y finalmente el componente celular (19%). Dentro de la colección de ESTs de yuca se identificaron 3.709 microsatélites que podrán ser empleados como marcadores moleculares. Este estudio representa una contribución importante al conocimiento de la estructura genómica funcional de la yuca y se constituye en una herramienta para la identificación de genes asociados a características de interés agrícola para posteriores programas de mejoramiento genético.


Cassava (Manihot esculenta) is the main source of calories for more than 1,000 millions of people around the world and has been consolidated as the fourth most important crop after rice, corn and wheat. Cassava is considered tolerant to abiotic and biotic stress conditions; nevertheless these characteristics are mainly present in non-commercial varieties. Genetic breeding strategies represent an alternative to introduce the desirable characteristics into commercial varieties. A fundamental step for accelerating the genetic breeding process in cassava requires the identification of genes associated to these characteristics. One rapid strategy for the identification of genes is the possibility to have a large collection of ESTs (Expressed Sequence Tag). In this study, a complete analysis of cassava ESTs was done. The cassava ESTs represent 80,459 sequences which were assembled in a set of 29,231 unique genes (unigen), comprising 10,945 contigs and 18,286 singletones. These 29,231 unique genes represent about 80% of the genes of the cassava’s genome. Between 5% and 10% of the unigenes of cassava not show similarity to any sequences present in the NCBI database and could be consider as cassava specific genes. A functional category was assigned to a group of sequences of the unigen set (29%) following the Gene Ontology vocabulary. The molecular function component was the best represented with 43% of the sequences, followed by the biological process component (38%) and finally the cellular component with 19%. In the cassava ESTs collection, 3,709 microsatellites were identified and they could be use as molecular markers. This study represents an important contribution to the knowledge of the functional genomic structure of cassava and constitutes an important tool for the identification of genes associated to agricultural characteristics of interest that could be employed in cassava breeding programs.

8.
Genet. mol. biol ; 33(4): 795-806, 2010. graf, tab
Article in English | LILACS | ID: lil-571541

ABSTRACT

Sequences potentially associated with coffee resistance to diseases were identified by in silico analyses using the database of the Brazilian Coffee Genome Project (BCGP). Keywords corresponding to plant resistance mechanisms to pathogens identified in the literature were used as baits for data mining. Expressed sequence tags (ESTs) related to each of these keywords were identified with tools available in the BCGP bioinformatics platform. A total of 11,300 ESTs were mined. These ESTs were clustered and formed 979 EST-contigs with similarities to chitinases, kinases, cytochrome P450 and nucleotide binding site-leucine rich repeat (NBS-LRR) proteins, as well as with proteins related to disease resistance, pathogenesis, hypersensitivity response (HR) and plant defense responses to diseases. The 140 EST-contigs identified through the keyword NBS-LRR were classified according to function. This classification allowed association of the predicted products of EST-contigs with biological processes, including host defense and apoptosis, and with molecular functions such as nucleotide binding and signal transducer activity. Fisher's exact test was used to examine the significance of differences in contig expression between libraries representing the responses to biotic stress challenges and other libraries from the BCGP. This analysis revealed seven contigs highly similar to catalase, chitinase, protein with a BURP domain and unknown proteins. The involvement of these coffee proteins in plant responses to disease is discussed.


Subject(s)
Humans , alpha 1-Antitrypsin , Information Science/statistics & numerical data , Mutation
9.
The Korean Journal of Parasitology ; : 103-107, 2009.
Article in English | WPRIM | ID: wpr-156347

ABSTRACT

The genus Acanthamoeba can cause severe infections such as granulomatous amebic encephalitis and amebic keratitis in humans. However, little genomic information of Acanthamoeba has been reported. Here, we constructed Acanthamoeba expressed sequence tags (EST) database (Acanthamoeba EST DB) derived from our 4 kinds of Acanthamoeba cDNA library. The Acanthamoeba EST DB contains 3,897 EST generated from amebae under various conditions of long term in vitro culture, mouse brain passage, or encystation, and downloaded data of Acanthamoeba from National Center for Biotechnology Information (NCBI) and Taxonomically Broad EST Database (TBestDB). The almost reported cDNA/genomic sequences of Acanthamoeba provide stand alone BLAST system with nucleotide (BLAST NT) and amino acid (BLAST AA) sequence database. In BLAST results, each gene links for the significant information including sequence data, gene orthology annotations, relevant references, and a BlastX result. This is the first attempt for construction of Acanthamoeba database with genes expressed in diverse conditions. These data were integrated into a database (http://www.amoeba.or.kr).


Subject(s)
Animals , Acanthamoeba/genetics , Databases, Genetic , Expressed Sequence Tags
10.
Genet. mol. biol ; 30(3,suppl): 917-930, 2007. ilus, tab
Article in English | LILACS | ID: lil-467270

ABSTRACT

In silico expression profiles, of the discovered 3,103 citrus ESTs putatively encoding for PR protein families (PR-1 to PR-17), were evaluated using the Brazil citrus genome EST CitEST/database. Hierarchical clustering was displayed to identify similarities in expression patterns among citrus PR-like gene families (PRlgf) in 33 selected cDNA libraries. In this way, PRlgf preferentially expressed by organ and citrus species, and library conditions were highlighted. Changes in expression profiles of clusters for each of the 17 PRlgf expressed in organs infected by pathogens or drought-stressed citrus species were displayed for relative suppression or induction gene expression in relation to the counterpart control. Overall, few PRlgf showed expression 2-fold higher in pathogen-infected than in uninfected organs, even though the differential expression profiles displayed have been quite diverse among studied species and organs. Furthermore, an insight into some contigs from four PRlgf pointed out putative members of multigene families. They appear to be evolutionarily conserved within citrus species and/or organ- or stress-specifically expressed. Our results represent a starting point regarding the extent of expression pattern differences underlying PRlgf expression and reveal genes that may prove to be useful in studies regarding biotechnological approaches or citrus resistance markers.

11.
Genet. mol. biol ; 30(3,suppl): 965-971, 2007. graf, tab
Article in English | LILACS | ID: lil-467274

ABSTRACT

The Citrus ESTs Sequencing Project (CitEST) conducted at Centro APTA Citros Sylvio Moreira/IAC has identified and catalogued ESTs representing a set of citrus genes expressed under relevant stress responses, including diseases such as citrus variegated chlorosis (CVC), caused by Xylella fastidiosa. All sweet orange (Citrus sinensis L. Osb.) varieties are susceptible to X. fastidiosa. On the other hand, mandarins (C. reticulata Blanco) are considered tolerant or resistant to the disease, although the bacterium can be sporadically detected within the trees, but no disease symptoms or economic losses are observed. To study their genetic responses to the presence of X. fastidiosa, we have compared EST libraries of leaf tissue of sweet orange Pêra IAC (highly susceptible cultivar to X. fastidiosa) and mandarin ‘Ponkan’ (tolerant) artificially infected with the bacterium. Using an in silico differential display, 172 genes were found to be significantly differentially expressed in such conditions. Sweet orange presented an increase in expression of photosynthesis related genes that could reveal a strategy to counterbalance a possible lower photosynthetic activity resulting from early effects of the bacterial colonization in affected plants. On the other hand, mandarin showed an active multi-component defense response against the bacterium similar to the non-host resistance pattern.

12.
Genet. mol. biol ; 30(3,suppl): 1009-1018, 2007. ilus, tab
Article in English | LILACS | ID: lil-467279

ABSTRACT

Nearly 65,000 citrus EST (Expressed Sequence Tags) have been investigated using the CitEST project database. Microsatellites were investigated in the unigene sequences from Citrus spp. and Poncirus trifoliata. From these sequences, approximately 35 percent of the non-redundant ESTs contained SSRs. The frequencies of different SSR motifs were similar between Citrus spp and trifoliate orange. In general, mononucleotide repeats appeared to be the most abundant SSRs in the CitEST database, but we also identify di-, tri-, tetra-, penta- and hexanucleotide repeats. The AG/CT and AAG/CTT were the most common dinucleotide and trinucleotide motifs, with frequencies of 54.4 percent and 25.2 percent, respectively. Primer sequences flanking SSR motifs were successfully designed and synthesized. After in silico polymorphism analysis, a subset of sixty-eight primers was validated in different Citrus spp. and Poncirus trifoliata. PCR-amplification revealed polymorphism in citrus with all tested primer pairs and showed the potential of these markers for linkage mapping. Our study showed that the CitEST database can be exploited for the development of SSR markers that can amplify Citrus spp. and related genus for comparative mapping and other genetic analyses.

13.
Genet. mol. biol ; 30(3,suppl): 1019-1023, 2007. tab
Article in English | LILACS | ID: lil-467280

ABSTRACT

In order to obtain a better understanding of what is citrus, 33 cDNA libraries were constructed from different citrus species and genera. Total RNA was extracted from fruits, leaves, flowers, bark, seeds and roots, and subjected or not to different biotic and abiotic stresses (pathogens and drought) and at several developmental stages. To identify putative promoter sequences, as well as molecular markers that could be useful for breeding programs, one shotgun library was prepared from sweet orange (Citrus sinensis var. Olimpia). In addition, EST libraries were also constructed for a citrus pathogen, the oomycete Phythophthora parasitica in either virulent or avirulent form. A total of 286,559 cDNA clones from citrus were sequenced from their 5’ end, generating 242,790 valid reads of citrus. A total of 9,504 sequences were produced in the shotgun library and the valid reads were assembled using CAP3. In this procedure, we obtained 1,131 contigs and 4,083 singletons. A total of 19,200 cDNA clones from P. parasitica were sequenced, resulting in 16,400 valid reads. The number of ESTs generated in this project is, to our knowledge, the largest citrus sequence database in the world.

14.
Genomics & Informatics ; : 86-93, 2005.
Article in English | WPRIM | ID: wpr-62312

ABSTRACT

Human brain EST data provide important clues for our understanding of the molecular biology associated with the function of the normal brain and the molecular pathophysiology with brain disorders. To systematically and efficiently study the function and disorders of the human brain, 45,773 human brain ESTs were collected from 27 human brain cDNA libraries, which were constructed from normal brains and brain disorders such as brain tumors, Parkinson's disease (PD) and epilepsy. An analysis of 45,773 human brain ESTs using our EST analysis pipeline resulted in 38,396 high-quality ESTs and 35,906 ESTs, which were coalesced into 8,246 unique gene clusters, showing a significant similarity to known genes in the human RefSeq, human mRNAs and UniGene database. In addition, among 8,246 gene clusters, 4,287 genes (52%) were found to contain full-length cDNA clones. To facilitate the extraction of useful information in collected these human brain ESTs, we developed a user-friendly interface system, the Korea Brain Unigene Database (KBUD). The KBUD web interface allows access to our human brain data through three major search modes, the BioCarta pathway, keywords and BLAST searches. Each result when viewed in KBUD offers comprehensive information concerning the analyzed human brain ESTs provided by our data as well as data linked to various other public databases. The user-friendly developed KBUD, the first world-wide web interface for human brain EST data with ESTs of human brain disorders as well as normal brains, will be a helpful system for developing a better understanding of the underlying mechanisms of the normal brain well as brain disorders. The KBUD system is freely accessible at http://kugi.kribb.re.kr/KU/cgi-bin/brain.pl.


Subject(s)
Humans , Brain Diseases , Brain Neoplasms , Brain , Clone Cells , DNA, Complementary , Epilepsy , Estrone , Expressed Sequence Tags , Gene Library , Korea , Molecular Biology , Multigene Family , Parkinson Disease , RNA, Messenger
15.
The Korean Journal of Parasitology ; : 149-156, 2005.
Article in English | WPRIM | ID: wpr-215235

ABSTRACT

Four hundred and sixty five randomly selected clones from a cDNA library of Blattella germanica were partially sequenced and searched using BLAST as a means of analyzing the transcribed sequences of its genome. A total of 363 expressed sequence tags (ESTs) were generated from 465 clones after editing and trimming the vector and ambiguous sequences. About 42% (154/363) of these clones showed significant homology with other data base registered genes. These new B. germanica genes constituted a broad range of transcripts distributed among ribosomal proteins, energy metabolism, allergens, proteases, protease inhibitors, enzymes, translation, cell signaling pathways, and proteins of unknown function. Eighty clones were not well-matched by database searches, and these represent new B. germanica-specific ESTs. Some genes which drew our attention are discussed. The information obtained increases our understanding of the B. germanica genome.


Subject(s)
Male , Female , Animals , Sequence Alignment , Reverse Transcriptase Polymerase Chain Reaction , Molecular Sequence Data , Expressed Sequence Tags , Blattellidae/genetics , Base Sequence
16.
Chinese Journal of Parasitology and Parasitic Diseases ; (6): 26-29, 2000.
Article in Chinese | WPRIM | ID: wpr-413196

ABSTRACT

[Objective] To obtain the genetic information on Necator americanus and to search for the purpose genes.[Methods] Mrna was isolated from the third stage larvae of Necator americanus maintained in hamsters.Double strand Cdna was synthesized and ligated to ΛzapⅡ vector to construct the Cdna library.Expresed se-quence tages (ESTs) were obtained by single pass sequencing of randomly isolated Cdna clones from the es-tablished library.[Results] A Cdna librazy of N.americanus was successfully constructed with high recombi-nant efficiency.The titer of unamplified library was 1×107.The insert size was about 750~3000bp.Of 11 ESTs obtained from the library,7 have a significant homology with certain functional genes.[Conclunsion]A high quality and high representative Cdna library of N.americanus was constructed at the first time and ome functional genes were identified from the library by ESTs.

17.
Chinese Journal of Parasitology and Parasitic Diseases ; (6)1987.
Article in Chinese | WPRIM | ID: wpr-584742

ABSTRACT

Subjective To acquire and analyze adult stage Schistosoma japonicum (Chinese strain) expressed sequence tags and new genes from an adult S. japonicum cDNA library, and to search new vaccine candidates and drug targets. Methods A cDNA library was constructed from adult stage S. japonicum. Clones were selected randomly from the cDNA library and were se-quenced. ESTs and new genes were acquired after analysis in GenBank databases by BLAST and other programs. All ESTs and new genes were submitted to GenBank and received accession numbers. Results 149 ESTs were acquired from a total 382 clones that were randomly selected from the adult S. japonicum cDNA library. All ESTs were successfully submitted to the dbEST at Genbank. Some of them were homologous with sequences of male, female, egg, schistosomula, cercaria and miracidia of S. japonicum. 18 new genes of adult S. japonicum were acquired. Some genes were housekeeping genes and some genes might be interesting as vaccine candidates or drugs targets. Conclusions The EST straltegy is a rapid, efficient and economical method to acquire ESTs and to discover new genes of adult stage S. japonicum from cDNA libraries.[

18.
Chinese Journal of Endocrinology and Metabolism ; (12)1986.
Article in Chinese | WPRIM | ID: wpr-674874

ABSTRACT

Objective To explore the characteristics of the gene expression in Cushing′s adrenal corticoadenoma and its associated molecular mechanism. Methods cDNA libraries of human normal adrenal and the adenoma were constructed. Large scale sequencing and bioinformatics were used. Results From the tumor library, 2089 expressed sequence tags (ESTs) were obtained with a successful rate of 98.9%. The functional distribution of the known genes in the tumor coincided with that in normal adrenal. Some synthases such as 3bHSDⅡ and P450c17A involved in steroidogenesis were expressed more highly in the adenoma than those in normal adrenal, especially 3bHSDⅡ. By comparison with the expression abundance of genes and statistic analysis, the differences were significant in 45 genes between the tumor and normal adrenal libraries. Interestingly, some genes associated with tumorigenesis, including heat shock protein 90, metallopanstimulin (MPS1), adenine nucleotide translocators etc, were highly expressed in the adenoma. However, those genes related to apoptosis and suppressing proliferation were not found in the tumor library. Conclusion The adenoma shows exuberant activities in steroid synthesis. Not only does the tumor highly express the genes associated with tumorigenesis, but also those related to apoptosis and suppressing proliferation lowly existed in the tumor.

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