%0 Journal Article
%T Construction of recombinant retroviral vector carrying porcine interferon-gamma and its expression in porcine kidney cells (PK-15)<br>携带猪γ干扰素基因逆转录病毒载体的构建及其在猪肾细胞(PK-15)中的表达
%A YAO Qing-xia
%A LIU Xin-wen
%A QIAN Ping
%A GUO Dong-chun
%A CHEN Huan-chun
%A <br>姚清侠
%A 刘新文
%A 钱平
%A 郭东春
%A 陈焕春
%J 微生物学报
%D 2007
%I 
%X Porcine interferon-gamma (PoIFN-gamma) of Chinese local brand, Meishan porcine, was cloned and inserted into retroviral vector pLXSN (neo r) . Using Lipofectamine, this recombinant plasmid was transfected into retroviral packing cell line, PA317 cells. These transfected cells were selected by DMEM containing 400microg/mL G418 for one week. RNA was extracted from the supernatant of these selected PA317 cells and the PoIFN-gamma gene could be amplified by RT-PCR. Pocine kidney cells and PK-15 cells were infected by the supernatant and were selected by 400 microg/mL, 600 microg/mL and 800 microg/mL G418, respectively. Those PK-15 cells were detected by indirect immunofluorescence assay and it was found that PoIFN-gamma mainly anchored in cellular membrane. The supernatant of the selected PK-15 was tested for the antiviral bioactivity after 48 hours of passage. The anti-VSV (vesicular stomatitis virus) activity in MDBK (bovine kidney cell) was 1200IU/10(6) cells. In addition, the effect of rPoIFNgamma-anti-FMDV was determined using cytopathic effect inhibition. The results indicate that PoIFN-gamma has been inserted into retroviral vector and recombinant retrovirus has been successfully packaged in PA317 cells. Furthermore, this retrovirus can infect PK-15 cells and express PoIFN-gamma with natural antiviral bioactivity and can inhibit VSV and FMDV.
%K Porcine interferon-gamma
%K Retroviral vector
%K PA317
%K PK-15
%K Expression
%K Antiviral activity
%K VSV
%K FMDV<br>猪γ干扰素
%K 逆转录病毒
%K PA317细胞
%K PK-15细胞
%K 表达
%K 抗病毒活性
%K 水泡性口炎病毒
%K 口蹄疫病毒
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=4CE92F63B53FD591&yid=A732AF04DDA03BB3&vid=F4B561950EE1D31A&iid=CA4FD0336C81A37A&sid=A8DE7703CC9E390F&eid=3986B25773CB6C30&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=15