Cloning And Expression Of ORF115a And ORF127 Gene From Helicoverpa Armigera Nucleocapsid Nucleopolyhedrorius

Helicoverpa armigera is a very important agricultural pest, harming to many crops, especially to cotton. Further more, it has serious resistance to many pesticides. While HeI icoverpa armigera single NPV, (HaSNPV) is the natural pathogeny microorganism to Helicoverpa armigera, can be used in biological control. With the development of molecule biology, the function of many genes have been understood, such as HaSNPV ORF128, HaSNPV ORF135 and HaSNPV ORF Ha122, but many genes' function is still unknown, such as HaSNPV ORF115a and HaSNPV ORF127. In this research, we analyze the biological information of HaSNPV ORF115a and HaSNPV ORF127 genes, clone and express them in E. coli. The results are the foundation for the futher research of gene function. The conclusion of this research is as follows:1 The result of the preliminary research of ORF115a from Heliocoverpa armigern nuclear polyhedrosis virusIn this research, we analyze the biological information of HaSNPV ORF115a, find that in HaSNPV-C1 genome Ha115a is located between 109,493bp and 110, 419bp ,and 926 nucleotide in coding section encodes a putative protein of 308 amino acid residues with a predicted molecular weight of 37.135KDa. The PI is 5.14. We Use ExPASy server to analyze the deduced protein, find that there is no signal petite, trans-membrance region, nucleus location region, but find some of important motifs, including one thiolases active site, one thiolases acyl-enzyme intermediate signature, one thiolases signature. The phylogenetic tree showed the amino acid sequence of ORF115a from Heliocoverpa armigern nuclear polyhedrosis virus and ORF119 from Helicoverpa zea nuclear polyhedrosis virus have high homology.With primers designed according to ORF115a from HaSNPV C1 geneone sequence (AF303045) published on GenBank, a DNA fragment of the HaSNPV was obtained by PCR amplification and was futher cloned into pMD18-T Easy vector. We get pMD18-T - ORF115a plasmid, and the plasmid was digested by BamH I and Xho I, we joint the fragment to pGEX-4T-2 vector, we get pGEX-4T-2- ORF115a , pGEX-4T-2- ORF115a plasmid was identified by BamH I and Xho I double enzyme digested, We transform the plasmid to BL21, according to SDS- PAGE...