ACCURATE INVITRO SPLICING OF HUMAN BETA-GLOBIN RNA
ACCURATE INVITRO SPLICING OF HUMAN BETA-GLOBIN RNA
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DOI：
10.1093/nar/10.18.5429
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发表时间：
1982-01-01
影响因子：
14.9
通讯作者：
WEISSMAN, SM
中科院分区：
文献类型：
--
作者：
KOLE, R;WEISSMAN, SM
从外源DNA模板转录的人β-珠蛋白RNA在HeLa细胞的浓缩全细胞提取物中体外拼接。用引物延伸技术，我们证明了小的插入序列是准确剪接的，剪接点上的产物序列与人网织红细胞β-珠蛋白mRNA的序列完全相同，剪接反应效率低。含有内含子和终止于多聚腺苷酸化位点上游的RNA转录物被最有效地剪接。终止于多聚腺苷酸化位点下游的转录物根本不被剪接。在小内含子中携带额外剪接位点的地中海贫血β-珠蛋白RNA也被剪接，尽管产量较低。
Human β-globin RNA transcribed from an exogenous DNA template is splicedin vitroby concentrated whole cell extracts from HeLa cells. Using the primer extension technique, we have shown that the small intervening sequence is spliced accurately and that the sequence of the product across the splice junction 1s identical to that of β-globin mRNA prepared from human reticulocytes.The efficiency of the splicing reaction is low. The RNA transcript containing both introns and terminated upstream from the polyadenylation site is spliced most efficiently. The transcript which is terminated downstream from the polyadenylation site is not spliced at all.Thalassemic β-globin RNA which carries an extra splice site 1n the small intron is also spliced, albeit with a low yield.