内含子大小对逆转录病毒转录本剪接效率的影响。
Effect of intron size on splicing efficiency in retroviral transcripts.
作者信息
Cullen B R, Kopchick J J, Stacey D W
出版信息
Nucleic Acids Res. 1982 Oct 11;10(19):6177-90. doi: 10.1093/nar/10.19.6177.
Abstract
To study the effect of intron size on splicing efficiency we have varied the size of the avian leukosis virus (ALV) env mRNA intron in a cloned ALV genome. This was accomplished by deletion of ALV sequences or insertion of phage lambda DNA. The effect of these modifications on splicing was analyzed by microinjection of the modified clones into RSV(-) chicken cells. Viral env mRNA when transcribed and properly spliced within these cells complemented the RSV(-) env deficiency leading to the production of focus forming units. Using this assay it was shown that deletion of up to 3.7 kb of the 4.68 kb env intron did not inhibit correct splicing nor did insertion of up to 8 kb of phage lambda DNA prevent splicing. Our results indicate that intron size can be varied over a wide range without preventing splicing.
摘要
为了研究内含子大小对剪接效率的影响，我们在克隆的禽白血病病毒（ALV）基因组中改变了禽白血病病毒env mRNA内含子的大小。这是通过删除ALV序列或插入噬菌体λ DNA来实现的。通过将修饰后的克隆显微注射到RSV（-）鸡细胞中，分析了这些修饰对剪接的影响。当病毒env mRNA在这些细胞中转录并正确剪接时，可弥补RSV（-）env缺陷，从而产生集落形成单位。利用该检测方法表明，删除4.68 kb env内含子中长达3.7 kb的片段不会抑制正确剪接，插入长达8 kb的噬菌体λ DNA也不会阻止剪接。我们的结果表明，内含子大小可以在很宽的范围内变化而不影响剪接。
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