Synthesis of predominantly unspliced cytoplasmic RNAs by chimeric herpes simplex virus type 1 thymidine kinase-human beta-globin genes.
Synthesis of predominantly unspliced cytoplasmic RNAs by chimeric herpes simplex virus type 1 thymidine kinase-human beta-globin genes.
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通过嵌合单纯疱疹病毒 1 型胸苷激酶-人 β-珠蛋白基因合成主要未剪接的细胞质 RNA。
DOI：
10.1128/mcb.5.8.1894-1900.1985
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发表时间：
1985
影响因子：
5.3
通讯作者：
Weissman,SM
中科院分区：
文献类型：
--
作者：
Greenspan,DS;Weissman,SM
单纯疱疹病毒1型胸苷激酶（tk）基因缺乏内含子，在没有剪接的情况下产生稳定的mRNA。我们将正常人β-珠蛋白基因的第一个外显子、第一个内含子（第一个介入序列，命名为IVS1）和第二外显子的大部分插入到该基因的3 '非翻译区，制备了一个杂交基因。尽管这个杂交基因包含了IVS1剪接所需的所有珠蛋白序列，但在长期和短期表达试验中，主要是产生了未剪接的稳定细胞质RNA。此外，无论内含子位于正义取向还是反义取向，都可以检测到稳定的未剪接的细胞质RNA。IVS1的有效剪接是通过删除大部分tk编码序列或通过将球蛋白序列从tk基因的3 ‘区重新定位到5 ’非翻译区来获得的。
The herpes simplex virus type 1 thymidine kinase (tk) gene lacks introns and produces stable mRNA in the absence of splicing. We have prepared a hybrid gene by placing the first exon, first intron (first intervening sequence, designated IVS1), and most of the second exon of the normal human β-globin gene into the 3′ untranslated region of thetkgene. Although this hybrid gene contains all globin sequences presumed necessary for the splicing of IVS1, predominantly, unspliced stable cytoplasmic RNA is produced in both long- and short-term expression assays. Moreover, stable unspliced cytoplasmic RNA is detected whether the intron is situated in a sense or an antisense orientation. Efficient splicing of IVS1 is obtained either by deleting the majority oftkcoding sequences or by relocating the globin sequences from the 3′ to the 5′ untranslated region of thetkgene.