Molecular clones representing Sendai virus genes P, NP and M.
Molecular clones representing Sendai virus genes P, NP and M.
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代表仙台病毒基因 P、NP 和 M 的分子克隆。
DOI：
10.1099/0022-1317-64-8-1679
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
Kingsbury,DW
中科院分区：
文献类型：
--
作者：
Gupta,KC;Morgan,EM;Kitchingman,G;Kingsbury,DW
仙台病毒基因 P、NP 和 M 片段的 DNA 拷贝是通过对从感染细胞中提取的病毒 mRNA 种类进行逆转录而获得的，并被克隆到质粒 pBR322 中。通过病毒 mRNA 的体外杂交抑制翻译来鉴定基因。 NP mRNA 的杂交选择证实了 NP 基因克隆的身份。该插入片段的部分测序表明它代表基因的 5' 末端区域，包含转录终止信号。 DNA 插入片段与电泳分离的变性 mRNA 种类的印迹杂交表明，P、NP 和 M 信息的大小分别为 2400、2100 和 1500 个核苷酸。先前在 NP 和 M 基因中鉴定出的特定 T1 核糖核酸酶抗性寡核苷酸通过与各自的插入片段杂交来选择。尽管大多数插入片段小于 500 个碱基对，仅占 P 基因的 16% 和 NP 基因的 24%，但其中一个包含 700 个碱基对的 M 基因插入片段几乎占该基因的一半。这些克隆的病毒基因片段将有助于进一步研究该模型副粘病毒的分子生物学。
DNA copies of segments of Sendai virus genes P, NP and M, obtained by reverse transcription of virus mRNA species extracted from infected cells, were cloned in plasmid pBR322. Genes were identified by hybrid-arrested translation of viral mRNAsin vitro. Hybrid selection of NP mRNA confirmed the identity of an NP gene clone. Partial sequencing of this insert showed that it represents the 5′-terminal region of the gene, containing transcription termination signals. Hybridization of DNA inserts to blots of electrophoretically separated, denatured mRNA species indicated that the P, NP and M messages had sizes of 2400, 2100 and 1500 nucleotides, respectively. Specific T1 ribonuclease-resistant oligonucleotides, previously identified in the NP and M genes, were selected by hybridization to the respective inserts. Although most of the inserts are smaller than 500 base pairs, representing no more than 16% of the P gene and 24% of the NP gene, one of the M gene inserts, comprising 700 base pairs, represents almost half of that gene. These cloned virus gene segments will assist further investigations of the molecular biology of this model paramyxovirus.