Genome-wide analysis of LBD(lateral organ boundaries domain) gene family in Cannabis sativa of traditional Chinese medicine hemp seed / 中国中药杂志

LBD(lateral organ boundaries)transcription factors play an important role in the regulation of plant growth, development and secondary metabolism. In order to explore the function of LBD genes in cannabis, the Cannabis sativa genome and transcriptome were used to identify the C. sativa LBD gene family, and analyzed their expression patterns. Our results showed that the cannabis LBD contains 32 members, which were divided into two major categories, seven sub-families. Class Ⅰ was divided into 5 sub-families, named Class Ⅰ_a to Class Ⅰ_e, while Class Ⅱ was divided into 2 sub-families, including Class Ⅱ_a and Class Ⅱ_b. Analysis showed that the number of amino acids encoded LBDs was between 172 and 356, and the isoelectric point was between 4.92 and 9.43. The mole-cular weight of LBD was between 18 862.92 Da and 40 081.33 Da, and most members are located in the nucleus. Chromosome positioning of LBD showed that 32 members were unevenly distributed on 10 chromosomes of C. sativa LBD transcription factor domain, gene structure and motifs are relatively conservative, and the characteristics of different class members are similar. The upstream promoter region of the gene contains a variety of cis-acting elements related to plant hormones and environmental factors, C. sativa LBD genes have different expression patterns in the stems, leaves, and flowers of ZYS varieties(low tetrahydrocannabinol, high cannabidiol). The members of the LBD gene family are mainly expressed in the flowers and stems of ZYS varieties, while members expressed in the leaves very few; Class Ⅱ members CsLBD21 and CsLBD23 are expressed in flowers and stems, and CsLBD8 and CsLBD18 are expressed in flowers, stems and leaves. These genes may participate in the growth and development of cannabis and affect the biosynthesis of cannabinoids. This study laid the foundation for the subsequently functional research of the cannabis LBD gene family.

Genome-wide Analysis of LBD (LATERAL ORGAN BOUNDARIES Domain) Gene Family in Brassica rapa

ABSTRACT LOB (lateral organ boundaries)-domain proteins define a family of plant-specific transcription factors involved in developmental process from embryogenesis to seed production. They play a crucial role in shaping the plant architecture through coordinating cell fate at meristem to organ boundaries. Identification of LBD genes from Brassica rapa genome, and analysis of phylogeny，gene structure, chromosome location, phylogenetic and tissue expression pattern analysis of LBD family genes in Chinese cabbage will be useful to the functions identification of plant LBD genes. Based on Brassica rapa genome database and bioinformatic method, Chinese cabbage LBD family genes were identified and the genes were sequenced. A phylogenetic tree was created using the MEGA5 program. Gene structure and chromosomes location were done by MapDraw, GSDS and Clustal X. Expression pattern of LBD genes at different development stages was analyzed based on RNA-seq. A total of 62 LBD genes were identified and could be classified into two classes and four subclasses according to the gene structure and conserved domain phylogeny relationship. Distribution mapping showed that the predicted LBDs were unevenly localized on all the 10 chromosomes, suggesting that they have an extensive distribution on the Brassica rapa chromosomes. Most of the LBD genes had differential expression pattern and showed highly diverse tissue-specific expression and functional diversity. To our knowledge, this is the first report of a genome wide analysis of the Brassica rapa LBD gene family, which would provide valuable information for understanding the classification and putative functions of the gene family.

Factors affecting digital image white balance of biological microscope / 医疗卫生装备

Objective To study the influencing factors of digital image white balance of the biological microscope and the way to obtain optimized image.Methods Color temperature meter was used to measure the influences of the factors and their correlations, and SASS software was employed for statistical analysis.Results Object lens might increase the color temperature by 200 to 300 K, LBD color filter could enhance it by 400 to 1 800 K. Reducing field stop by 90% might decrease color temperature by 400 K. ND6 filter, ND25 filter and aperture diaphragm had no influences on color temperature.Conclusion The electric and optical parameters of biological microscope have to be adjusted to gain proper white balance for digital image.

Cryoprotection effectiveness of low concentrations of natural and lyophilized LDL (low density lipoproteins) on canine spermatozoa / Eficiência de baixas concentrações de lipoproteínas de baixa densidade natural e liofilizada na crioproteto de espermatozoides caninos

The aim of this study was to evaluate the use of low concentrations of natural and lyophilized low density lipoprotein (LDL) from hen's egg yolk for cryopreservation of canine semen. Different ammonium sulphate concentrations were tested to extract LDL from egg yolk. The yolk was centrifuged, and LDL was isolated using 10, 20, 40, 45, or 50 percent ammonium sulphate solution (ASS). The LDL-rich floating fraction was collected for chemical characterization. Dry matter content was lowest (P<0.05) in the LDL extracted with the 50 percent ASS. The purification of LDL increased in association with increasing ammonium sulphate concentrations. SDS-PAGE showed that the 50 percent ASS solution yielded a purer fraction of LDL from egg yolk. For semen cryopreservation, TRIS extender was used replacing 20 percent egg yolk (control) by natural or lyophilized LDL using 1, 2, and 3 percent (w/v). Semen was centrifuged (755Xg for 7 min), diluted with one of the extenders, packed into 0.5mL straws (100x106 sperm/mL), and placed in a programmable cryopreservation machine. Thawed semen (37°C/ 30s) was analyzed for sperm motility, morphology, and by the hypoosmotic and epifluorescence tests (CFDA/ PI). Natural LDL extracted with 50 percent ASS was as effective as whole egg yolk to preserve canine frozen sperm when using low concentrations. The lyophilized LDL, mainly in the two higher concentrations tested (2 and 3 percent), was unsuitable to maintain the effectiveness of the LDL cryoprotective effect on dog sperm...

O objetivo deste estudo foi avaliar o uso de baixas concentrações da lipoproteína de baixa densidade (LBD) extraída da gema do ovo de galinha, nas formas natural e liofilizada, na criopreservado do sêmen canino. Diferentes concentrações de sulfato de amônio também foram testadas na extrato da LBD da gema do ovo. A gema foi centrifugada, sendo a LBD isolada usando-se soluto saturada de sulfato de amônio (SSA) nas concentrações de 10, 20, 40, 45 e 50 por cento. A frado rica em LBD foi coletada para caracterizado química. O conteœdo de matéria seca foi menor (P<0,05) na LBD extraída com SSA 50 por cento. A pureza da LBD melhorou medida que se aumentou a concentrado de SSA utilizada. SDS-PAGE mostrou que a SSA 50 por cento produziu uma frado mais pura de LBD oriunda da gema do ovo. Para o congelamento de sêmen, o meio diluidor TRIS teve a gema do ovo a 20 por cento (controle) substituída pela LBD a 1, 2 e 3 por cento (p/v), nas formas natural e liofilizada. O sêmen foi centrifugado (755xg por 7min), diluído em um dos meios diluidores em teste e envasado em palhetas de 0,5mL (100x106 sptz/mL), sendo congelado em máquina de congelamento programável. O sêmen descongelado (37°C/30s) foi analisado quanto motilidade e morfologia espermática e nos testes hiposm-tico e de epifluorescência (DACF/IP). A LBD natural extraída com SSA 50 por cento foi tão eficiente quanto a gema do ovo na preservado do espermatozoide canino congelado nas baixas concentrações testadas. A LBD liofilizada, principalmente as duas maiores concentrações (2 e 3 por cento), não foi adequada para manter o efeito crioprotetor da LBD sobre o espermatozoide canino...

Research of drug screening model based on recombinant hPPAR?-LBD as a target protein for lipid metabolism / 重庆医科大学学报

Objective:To establish a method of antitumor drug screening model based on recombinant hPPAR?-LBD as a target protein for lipid metabolism. Methods:pReceiver-B01-PPAR?- LBD expression plasmid was constructed and transferred into Escherichia coli BL2(1DE3)competent cells. Growth conditionswere optimized to induce soluble expression of hPPAR?-LBD recombinant protein. Purification was carried out by Nickel affinity chromatography. Ligand binding activity of the recombinant protein was determined by novel size exclusion chromatography and high performance liquid chromatography(SEC-HPLC)method with rosiglitazone as reference ligand and GW9662 as antagonist. Results:Under optimal conditions (16℃,0.6 mmol/L IPTG and induction time 20 h),the soluble recombinant protein was expressed successfully. After one-step purification with Nickel affinity chromatography,41 mg of recombinant protein with more than 95% purity could be obtained from per liter Luria-Bertani(LB)medium. The Kd and percentage ofspecific bindingofrosiglitazone and tetrazanbigen tohPPAR?-LBDare 625 nmol/L,65% and 1 000 nmol/L,60%,respectively.Conclusion:In this study,a fast,stable and simple method could be used successfully to screen antitumor drug.