Cloning Of Bombyx Mori Inhibitor Of Apoptosis Proteins Gene And Its Anti-apoptosis Effect

Apoptosis means gene under the control of cell death in the initiative known as programmed cell death (programmed cell death, PCD). However, inappropriate apoptosis, such as large number of apoptotic cells or apoptosis will not lead to the occurrence of diseases. IAPs were discovered in recent years for a new kind of cell apoptosis inhibitory protein, which is widely in many species. molecular mechanisms of apoptosis is a very important and conservative protein family. So far, two baculovirus IAP, seven mammalsIAP,. three Arthropod IAP and Drosophila IAP were discovered.But few studies Bombyx mori IAP.Under the silkworm genome sequence lap specific primers were designed. Iap gene which was amplified fragments using RT-PCR method was ligated into pUC18 vector for sequencing. The result showed that the genetic reading frame was 1041 bp in size, encoded 346 amino acids.PET28a-iap prokaryotic expression vector was constructed. Product of lap gene in E. coil was tested using SDS-PAGE and Western Blotting. a molecular weight of 38.8 kD band of specific expression was detected, which was line with the theoretical value. The result showed that IAP was expressec in E.coil. Quantitative PCR method was used to detect iap gene expression in UV stimulated silkworm cells and larvae. The results showed that iap gene expression increased with the dose of stimulation, and then fell over, declined to speculate because of excessive ultraviolet stimulate cell injury; on the other hand, the iapgene stress is the early gene expression.The recombinant virus r-iap infected Bm cell using baculovirus expression system, cell apoptosis was studyed by cell morphology and trypan blue staining method combining cell survival. The results indicated that the Silkworm IAP played host cell extended life and life cycle role.The correct pCDNA3.1-gfp-iap mammalian cells transfected into COS-7 and was higher expression. It was discussed that how iap gene played role in nerve cells in the treatment of nerves diseases.