van Strien E A, Zuidema D, Goldbach R W, Vlak J M
Department of Virology, Wageningen Agricultural University, The Netherlands.
J Gen Virol. 1992 Nov;73 ( Pt 11):2813-21. doi: 10.1099/0022-1317-73-11-2813.
The nucleotide sequence of a 1.1 kbp fragment of the multiple nucleocapsid nuclear polyhedrosis virus (MNPV) of Spodoptera exigua (Se) containing the polyhedrin gene was determined. An open reading frame (ORF) of 738 nucleotides (nt) was detected. This ORF encoded a protein of 246 amino acids with a predicted M(r) of 29K. The nucleotide and amino acid sequences were compared with the sequences of eight other NPV polyhedrins. The SeMNPV polyhedrin protein was most closely related to S. frugiperda MNPV polyhedrin with differences in only five amino acids, and most distantly related to the Lymantria dispar MNPV polyhedrin. The size of the mRNA was approximately 1,000 nt, as determined by Northern blot analysis. Using primer extension assays and S1 nuclease mapping the transcriptional start and stop sites of the polyhedrin mRNA were located. The 5' regulatory sequence appeared to be 44 nt in length with the mRNA start site predominantly at the first A of the TAAG consensus start sequence. Two degenerate poly(A) signals were found immediately downstream of the translational stop signal. The transcriptional stop was located approximately 230 nt downstream from the translational stop signal, in an AT-rich sequence that appears to be common to all baculovirus polyhedrin genes. The SeMNPV polyhedrin mRNA does not appear to be polyadenylated.
测定了甜菜夜蛾多核衣壳核型多角体病毒(SeMNPV)含多角体蛋白基因的1.1kbp片段的核苷酸序列。检测到一个738个核苷酸(nt)的开放阅读框(ORF)。该ORF编码一个246个氨基酸的蛋白质,预测分子量(M(r))为29K。将该核苷酸和氨基酸序列与其他8种NPV多角体蛋白的序列进行了比较。SeMNPV多角体蛋白与草地贪夜蛾MNPV多角体蛋白关系最为密切,仅5个氨基酸存在差异,与舞毒蛾MNPV多角体蛋白关系最远。通过Northern印迹分析确定,mRNA的大小约为1000nt。利用引物延伸试验和S1核酸酶作图定位了多角体蛋白mRNA的转录起始和终止位点。5'调控序列长度似乎为44nt,mRNA起始位点主要位于TAAG共有起始序列的第一个A处。在翻译终止信号下游立即发现两个简并的聚腺苷酸化信号。转录终止位于翻译终止信号下游约230nt处,在一个富含AT的序列中,该序列似乎是所有杆状病毒多角体蛋白基因所共有的。SeMNPV多角体蛋白mRNA似乎未进行聚腺苷酸化。