腺病毒2型信使核糖核酸形成过程中的体外核糖核酸-核糖核酸剪接
In vitro RNA-RNA splicing in adenovirus 2 mRNA formation.
作者信息
Blanchard J M, Weber J, Jelinek W, Darnell J E
出版信息
Proc Natl Acad Sci U S A. 1978 Nov;75(11):5344-8. doi: 10.1073/pnas.75.11.5344.
Abstract
"Splicing" of the precursor to an adenovirus mRNA was accomplished in isolated cell-free extracts. Nuclei were prepared from hypotonically swollen cells that had been labeled with [3H]uridine for 10 min prior to nuclear isolation. Addition of a "cytoplasmic" fraction was required for the splicing to occur. The nuclear precursor, a poly(A)-terminated RNA molecule approximately 5 kilobases long, contained sequences complementary to the 58.5--75.9 region of the adenovirus 2 genome, including those sequences spliced out of the mature mRNA molecule. The in vitro spliced product was a poly(A)-terminated RNA molecule identical in size to the cytoplasmic 72,000 Mr protein mRNA (2 kilobases long) in which the sequences encoded in the 70.7--75.9 region of the viral genome were spliced to those encoded at 58.7--65.6, with the sequences encoded at 66.1--70.7 deleted.
摘要
腺病毒信使核糖核酸（mRNA）前体的“剪接”在分离的无细胞提取物中完成。细胞核是从低渗肿胀的细胞中制备的，这些细胞在细胞核分离前用[³H]尿苷标记了10分钟。剪接的发生需要添加一种“细胞质”组分。核前体是一个约5千碱基长的聚腺苷酸化（poly(A)）终止的RNA分子，它包含与腺病毒2基因组58.5 - 75.9区域互补的序列，包括那些从成熟mRNA分子中剪接出去的序列。体外剪接产物是一个聚腺苷酸化终止的RNA分子，其大小与细胞质中72,000道尔顿（Mr）蛋白质的mRNA（2千碱基长）相同，其中病毒基因组70.7 - 75.9区域编码的序列与58.7 - 65.6区域编码的序列进行了剪接，而66.1 - 70.7区域编码的序列被删除。
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