TAZ overexpression is associated with epithelial-mesenchymal transition in cisplatin-resistant gastric cancer cells.

Gastric cancer is one of the common malignant diseases. The poor treatment outcome is mainly due to chemotherapeutic resistance. Therefore, it is important to determine the molecular mechanism of drug resistance in gastric cancer. To explore the mechanisms of cisplatin resistance in gastric cancer cells, several approaches were performed including MTT assay, real-time RT-PCR, western blot analysis, migration and invasion assays, wound healing assay, and transfection. We found that cisplatin-resistant (CR) gastric cancer cells acquired epithelial-mesenchymal transition (EMT) phenotype. The CR cells with EMT features obtained higher migratory and invasive activities. Moreover, we observed that TAZ was highly expressed in CR cells. Consistently, depletion of TAZ caused partial reversal of EMT to MET in CR cells. Our results suggest that TAZ plays a pivotal role in CR-induced EMT. Targeting TAZ could be a potential therapeutic strategy for gastric cancer.

[Sequencing analysis of the control region and intron1 region and characterization of two novel microsatellites in obese gene of pig].

For the first time, a 16 kb fragment of the porcine Ob gene, namely intron1, exon1, 5' region of Ob gene, was restrictively analyzed and sequenced by the primers designed in the portion of the swine Ob sequence. The first small 38 bp untranslated exon1 is located 11.1 kb upstream of the initiator ATG codon, and two novel microsatellites SW200 and SW160 are found in intron1. Promoter analysis of several putative binding sites revealed that this 300 bp promoter located at -1 to -300, including C/EBP and two Sp1, may be as effective as the longer promoter in directing leptin transcription. To examine microsatellites association with important economic traits, we statistically analyzed genotypes and alleles of the two microsatellites. Statistical analysis carried out by SAS 8.2 revealed significant positive correlation between the two microsatellites genotypes and the litter size in first parity of Erhualian.

Isolation and characterization of 70 novel microsatellite markers from ostrich (Struthio camelus) genome.

Microsatellite markers are widely used in linkage mapping, parentage testing, population genetic studies, and molecular evolution studies in many agricultural species, while only a limited number of ostrich (Struthio camelus) microsatellites have been isolated. Thus, we constructed a random small-insert genomic library and a microsatellite-enriched library containing CA repeats. Fourteen clones containing CA repeats were isolated from 3462 clones in the non-enriched library by radioactive screening and 248 positive clones were isolated from 300 sequenced clones from the enriched library by PCR screening. After the enrichment procedures, the proportion of clones containing CA repeats was raised to 78.8%, compared with 0.4% in the non-enriched libraries, indicating that the enrichment value approaches 200 fold, which decreased the time and cost of cloning. The number of complete simple CA repeats in these positive clones ranged from 5 to 29. The primers for 94 of these microsatellites were developed and used to detect polymorphisms, of which 61 loci exhibited length polymorphisms in 17 unrelated ostrich individuals. The new polymorphic microsatellite markers we have identified and characterized will contribute to the ostrich genetic map, parentage testing, and comparative genomics between avian species.

[Preliminary analysis on China Agricultural University chicken resource population based on genomic scanning].

Combining the technique of multiplex-PCR and the fluorescent semi-automated detection, a large-scale genome scanning was performed for 440 chickens, which was derived from China Agricultural University chicken resource families, within three generations. Fifty-five microsatellite markers were analyzed for this study. Those 55 microsatellite loci accorded with the characters of Mendelian co-inheritance. The heterozygosities ranged from zero to 0.89, with 72% of loci having a heterozygosity of more than 0.60. The polymorphism information content (PIC) ranged from 0 to 0.85, in which 70% of those loci had a PIC of more than 0.50 but their distribution varied in line A and line C. The allele frequency was significantly different between line A and line C at most loci (P < 0.01). At the same time, gene accordance inclination was found in line C. The Nei population resemble coefficient and standard genetic distance were 0.1002 and 0.8928.

[The factors affecting the efficiency of mutiplex PCR].

In the research,the outputs of different cycle parameters, PCR buffer and reaction volumes are compared. The results indicated that the annealing temperature, annealing time, elongated time and ingredient of PCR buffer affected mutiplex PCR, but the reaction volume and cycle number had few effect on it.

[Analysis of succeed percentage on sequencing ESTs and construction of porcine muscle cDNA library].

Using Longissimus Dorsi muscle as material and Lambda ZAP II as Vector, Xiang Pig Longissimus Dorsi muscle cDNA library has been constructed in our study. The results showed that the titration of the library was 3.4 x 10(7) pfu/ml, the recombinant percentage was 94%, and the fragment length of inserted average cDNA were 1.5 kb. The study pointed out that the more than 30 T insertion is the major factor for low percentage if sequencing the 3'-end.