Construction Of Antheraea Pernyi Nuclear Polyhedrosis Virus Double Promoter Transfer Vector Contains Genes Of IGF-1 And GFP

Insulin-like growth factors are a series of multi-functional cell proliferation regulatoryfactor. IGF-1 is a member of the super family which is a 7.65kD, single-strand, peptide-likegrowth factor consisting of 70 amino acid residues. It has several functions that are similarwith insulin, so it has an important practical value in the treatment of many diseases and has apromising perspective. Inartificial IGF-1 is little, expensive and instability. Baculovirusexpression vector system (BEVS) has been a powerful tool to investigate and produce severalof prokaryotic and eukaryotic proteins since it came out in 1980s, however recombined viruswas difficult to filtrate.The aim of this experiment is to construct the double-promoter transfer vector containsgenes of IGF-1 and GFP. Firstly, we have constructed the expressive vector IE1/pGFP-N2,the promoter of Antheraea pernyi Nuclear Polyhedrosis Virus IE-1 gene was cloned, and thensequencing was analyzed. The IE-1 promoter was inserted into vector pGFP-N2 Greenfluorescent protein. After transfectional examination, the expressive vector IE-1/pGFP-N2was successfully constructed.Secondly, we constructed colon vector consisting of the end sequence SV40 polyA,colon vector of IGF-1, and constructed colon vector GFP consisting of promoter IE-1 gene.IGF-1 is a peptide, consisting of 70 amino acid residues. In order to improve the expressionsof IGF-1, we constructed IGF-1 in series the colon vector.Finally, the aim of gene and reported gene was constructed in the vector of pApM. Thesequence is IGF-1-SV40 polyA-Pie-GFP. The excellently characteristic of vector have doublepromoter. At obtained the recombine virus, the cells consisting of recombine virus was visualgreen in fluorescence microscope, have evidently select marker, and improved the work offilter.