SV40晚期mRNA在来自CV-1细胞的分离细胞核中的体外剪接。
In vitro splicing of SV40 late mRNA in isolated nuclei from CV-1 cells.
作者信息
Hamada H, Igarashi T, Muramatsu M
出版信息
Nucleic Acids Res. 1980 Feb 11;8(3):587-99. doi: 10.1093/nar/8.3.587.
An in vitro splicing system utilizing isolated nuclei of SV40 infected cells has been developed. Nuclei were isolated from CV-1 cells at a late stage of SV40 infection after a pulse-labeling with 3H-uridine. In nuclei prepared under mild isotonic conditions, 19S viral coded RNA synthesized in vivo was converted in vitro into 16S mRNA. In contrast, the nuclei prepared with RSB, a hypotonic medium, showed a very low splicing activity only. Addition of a "nuclear extract" to these nuclei restored the activity almost to the original level. These results indicate that 1) 19S RNA is indeed a precursor to 16S mRNA 2) the splicing of 19S RNA into 16S RNA takes place in the nucleus, and 3) at least a part of the enzyme system required for splicing could be extracted from the nucleus. This in vitro system may be useful for the assay of the splicing enzyme(s).
一种利用SV40感染细胞的分离细胞核的体外剪接系统已被开发出来。在用3H-尿苷脉冲标记后，从SV40感染后期的CV-1细胞中分离细胞核。在轻度等渗条件下制备的细胞核中，体内合成的19S病毒编码RNA在体外被转化为16S mRNA。相比之下，用低渗介质RSB制备的细胞核仅表现出非常低的剪接活性。向这些细胞核中添加“核提取物”可将活性几乎恢复到原始水平。这些结果表明：1）19S RNA确实是16S mRNA的前体；2）19S RNA剪接成16S RNA发生在细胞核中；3）剪接所需的酶系统至少有一部分可以从细胞核中提取出来。这种体外系统可能有助于剪接酶的测定。
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