Pulford D J, Britton P
A.F.R.C., Institute for Animal Health, Compton Laboratory, Berkshire, U.K.
Virus Res. 1991 Mar;18(2-3):203-17. doi: 10.1016/0168-1702(91)90019-r.
Porcine transmissible gastroenteritis virus (TGEV) nucleoprotein and integral membrane protein genes were cloned into the vaccinia virus insertion vector, pGS20, in the correct orientation for expression under the control of the vaccinia P7.5K promoter. Recombinant vaccinia viruses were generated by in vivo homologous recombination of the insertion vector with the WR strain of vaccinia virus. Nucleoprotein (N) expressed by both recombinant vaccinia virus and TGEV had a relative molecular mass (Mr) of 47,000 and was susceptible to degradation at the C-terminus yielding discrete breakdown products. The integral membrane protein (M) expressed by a recombinant vaccinia virus and TGEV was sensitive to endoglycosidase H reducing the mature polypeptide of Mr 29,000 to a species of Mr 27,000. Expression of M by recombinant vaccinia virus was inhibited during early infection due to a cryptic vaccinia virus transcriptional termination signal within the TGEV coding sequence. Indirect immunofluorescence showed that both N and M were only localised in the cell cytoplasm of either TGEV or recombinant vaccinia virus immunoprecipitated specific TGEV antigens from lysates of TGEV infected cells but had little significant TGEV neutralising activity in vitro.
猪传染性胃肠炎病毒（TGEV）核蛋白基因和整合膜蛋白基因被以正确的方向克隆到痘苗病毒插入载体pGS20中，以便在痘苗P7.5K启动子的控制下表达。通过插入载体与痘苗病毒WR株进行体内同源重组产生重组痘苗病毒。重组痘苗病毒和TGEV表达的核蛋白（N）的相对分子质量（Mr）为47,000，并且在C末端易降解，产生离散的降解产物。重组痘苗病毒和TGEV表达的整合膜蛋白（M）对内切糖苷酶H敏感，使Mr 29,000的成熟多肽降解为Mr 27,000的一种多肽。由于TGEV编码序列内有一个隐蔽的痘苗病毒转录终止信号，重组痘苗病毒在早期感染期间对M的表达有抑制作用。间接免疫荧光显示，N和M都只定位于TGEV或重组痘苗病毒的细胞质中，从TGEV感染细胞的裂解物中免疫沉淀特异性TGEV抗原，但在体外几乎没有显著的TGEV中和活性。
