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Tung Looper Nuclear Polyhedrosis Virus P35 Gene

Posted on:2006-07-24Degree:MasterType:Thesis
Country:ChinaCandidate:Y J TaoFull Text:PDF
GTID:2193360155474520Subject:Microbiology
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In this paper, the function of Buzura suppressaria nuclear polyhedrosis virus (BsNPV) p35 was studied. A 940bp fragment was produced by PCR from BsNPV DNA and sequenced after cloned into plasmid pMD-18.We have cloned and expressed the gene in E.Coli cells and got high level yielding of protein. The protein have been assayed by the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) .The results showed, the molecular weight of this protein was about 62kD because this was a fusion protein constituted by P35 and GST protein.Next, BsNPVp35 was cloned into the MCS of pIZ/V5-His before its stop- codon was mutated and EGFP gene was cloned downstream of p35 gene, be sure that they are in the same reading frame. This recombinant vector was named pIZ/V5-p35-egfp. After it was transformed into HZ-AM1 cells, some apoptosis factors were added in order to investigate weather P35 can delay apoptosis in insect cells.Because p35 is a host-range limiting factor for baculovirus infections, some baculovirus' p35 protein have no anti-apoptosis or its function depended on the particularity of host, the results need to be analysised.
Keywords/Search Tags:Buzura suppressaria nuclear polyhedrosis virus, p35 gene, apoptosis, EGFP, Heliothis armigera, Cell culture
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