来自英国一株强毒力传染性胃肠炎病毒野毒株的核蛋白基因序列及其在酿酒酵母中的表达
Sequence of the nucleoprotein gene from a virulent British field isolate of transmissible gastroenteritis virus and its expression in Saccharomyces cerevisiae.
作者信息
Britton P, Cármenes R S, Page K W, Garwes D J, Parral F
机构信息
Division of Microbiology, A.F.R.C. Institute for Animal Disease Research, Compton Laboratory, Compton, nr. Newbury, Bertshire, RG16 ONN, UK.
出版信息
Mol Microbiol. 1988 Jan;2(1):89-99. doi: 10.1111/j.1365-2958.1988.tb00010.x.
Subgenomic mRNA from a virulent isolate of porcine transmissible gastroenteritis virus (TGEV) was used to produce cDNA which was sequenced. Two non-overlapping open reading frames (ORFs) were identified. The largest, encoding a polypeptide of 382 amino acids (relative molecular mass (M ) 43 483), was shown to be the viral nucleoprotein gene. The second ORF, found 3'to the larger ORF, encodes a polypeptide of 78 amino acids (M 9068) which has yet to be assigned to a viral product. The nucleoprotein gene was expressed in yeast cells under the control of two types of yeast promoters: the constitutive PGK promoter, and the inducible GAL1 promoter. Yeast cells containing recombinant plasmids, with the nucleoprotein gene in the correct orientation, produced a polypeptide of M, 47000, identical to the viral product, that reacted with a specific monoclonal antibody.
来自猪传染性胃肠炎病毒（TGEV）强毒株的亚基因组mRNA被用于制备cDNA并进行测序。鉴定出两个不重叠的开放阅读框（ORF）。其中最大的一个编码一个由382个氨基酸组成的多肽（相对分子质量（M）为43483），被证明是病毒核蛋白基因。第二个ORF位于较大ORF的3'端，编码一个由78个氨基酸组成的多肽（M为9068），其功能尚未确定为病毒产物。核蛋白基因在两种酵母启动子的控制下在酵母细胞中表达：组成型PGK启动子和诱导型GAL1启动子。含有重组质粒且核蛋白基因方向正确的酵母细胞产生了一个M为47000的多肽，与病毒产物相同，并且能与一种特异性单克隆抗体发生反应。
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