ABSTRACT
To clone overexpressed gene from human gastric carcinoma cell SGC-7901, DDRT-PCR technique is used with human gastric epithelial cell GES-1 as control. After cloned into pGEM -T vector, YA61, one of the overexpressed genes, was analyzed by dot blot and was sequenced then. The sequence gotten was then compared to GenBank data and analyzed by NCBI ORF Finder. Dot blot results showed that the gene YA61 was overexpressed in human gastric carcinoma cell SGC-7901. NCBI's sequence similarity search indicated that the gene YA61 was a new gene sequence. Open reading frame analysis demonstrated that the gene YA61 had one complete open reading frame. In conclusion, the gene YA61 was a new gene sequence that was overexpressed in human gastric carcinoma cell SGC-7901.
ABSTRACT
To look for new genes from human brain, get a fragment was obtained using adaptor primer and 3' anchor polymerase chain reaction (PCR) with the human adult whole brain cDNA as template. The fragment was cloned into T easy vector and automatically sequenced with 310 Genetic Analyzer. Later the whole length cDNA of this novel gene was got with the method of 3'rapid amplification of cDNA end (RACE). The whole length of cDNA of this novel gene is 2 024 bp. Chromsome location is at 14q11.2 including 16 extrons and 15 introns. After scanning the sequence against GenBank it is proved that the sequence is a new one. ORF analysis showed that there is a complete coding region in it,it can interprate a protein containing 357 amino acid residules. ProDom analysis result showed that there is an acyl carrier protein (ACP) like domain in it. The gene was banked into GenBank. Then, a pare of primers were designed and were used to amplify the coding region and cloned into pGEX-4T1 expressing vector to express it in E. coli . The Dot blotting and Northern blot showed that this novel gene is highly expressed in the normal adult human brain.
ABSTRACT
Aim To express the fusion protein hTNFα -hPgnK5 in E.Coli and to identify its activity. Methods The hTNFα -hPgnK5 gene express vector was constructed and the fusion protein was expressed in E.coli. The activity of fusion protein to inhibit the proliferation of tumor cells or angioendothelial cells in vitro was determined by MTT. Results Fusion protein hTNFα -hPgnK5 possessed immunoactivity in Western blot analysis and could inhibit proliferation of tumor cells or angioendothelial cells in vitro test. Conclusion The hTNFα gene and hPgnk5 gene can be coexpressed in prokaryotic system and expression products can inhibit the proliferation of tumor cells or angioendothelial cells in vitro.