The Investigation, Characterization Of Promoting Efficacy Of PERV And Construction Of Retroviral Vector

As we all know, donor organs are most deficient in clinic. People are fixing their eyes on xenotransplantation, and porcines are considered to be the most suitable donor up to now. However, PERV, integrating into the genome of porcines by the form of provirol DNA, could infect human-derived cells in vitro. As a result, the safety of xenotransplantation was on the agenda. Although China is rich in minipig resources and China carried out a lot of trials on xenotransplantation from porcines to human beings, the infectivity, expression and biological characteristic of PERV in pigs are rarely reported. As a result, it is quite necessary to understand the PERV existence situation, privide safe and reliable organs for transplantation, to understand the characterization of promoting efficacy of the PERV 5' untranslated region for further PERV biological characteristics. In addition, traditional retroviral vectors have some problems on low titre pseudotyping virions and biological safety while its transgene expression, but retroviral vector based on PERV may solve these problems. It is because that PERV exsist in the porcine genome and evolove with pig for hundreds of millions of years, and the copy numbers of full-length PERV sequences per genome (approximately 8 to 15) vary among swine strains, but any infection or disease due to PERV has not been detected, and because of the homologous sequence exsisting in procine cells, we could get high titre pseudotyping virions after tranfected the PERV retrovial vector into procine cells. Many researches focused on the evaluation of biological safety with porcine endogenous retrovirus in xenotransplantation, not a research in the retroviral vector based on 5'LTR of PERV, so our research work relate to construct PERV retroviral vector. Our research work includes the following:1. Existance status of PERV in Chinese Wuzhishan PigsWe conducted this research to investigate the existence and expression status of porcine endogenous retrovirus (PERV) in Chinese wuzhishan pigs. Based on PCR and RT-PCR methods established before in our lab. we employed universal env genotyping methods to detect the existence and expression status of PERV env-A,...