Blumberg B M, Giorgi C, Rose K, Kolakofsky D
J Gen Virol. 1984 Apr;65 ( Pt 4):769-79. doi: 10.1099/0022-1317-65-4-769.
A procedure is presented for isolating the nucleocapsid proteins, N and NP from vesicular stomatitis virus and Sendai virus respectively, in soluble form. These proteins were suitable for the determination of their blocked amino-terminal peptide sequences by gas-liquid chromatography/mass spectrometry at the low nanomole level. The N protein prepared by this procedure was previously shown to retain some of its expected biological activity. The sequence of 626 nucleotides from the 3' end of the Sendai virus genome, which includes the first one-third of the NP gene, was determined. Using this information, primer extension studies on intracellular Sendai virus mRNAs allowed the determination of the structure of the leader-NP intervening sequence and the 5' end of the NP mRNA. Comparison of the amino termini of the nucleocapsid proteins with their respective mRNA sequences revealed that these proteins are similarly processed in vivo.
本文介绍了一种分别从水疱性口炎病毒和仙台病毒中以可溶形式分离核衣壳蛋白N和NP的方法。这些蛋白适用于通过气液色谱/质谱法在低纳摩尔水平测定其封闭的氨基末端肽序列。通过该方法制备的N蛋白先前已证明保留了其一些预期的生物学活性。测定了仙台病毒基因组3′端626个核苷酸的序列，其中包括NP基因的前三分之一。利用这些信息，对细胞内仙台病毒mRNA进行引物延伸研究，从而确定前导-NP间隔序列的结构和NP mRNA的5′端。将核衣壳蛋白的氨基末端与其各自的mRNA序列进行比较，结果表明这些蛋白在体内的加工方式相似。
