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纯化的DNA在显微注射到非洲爪蟾卵母细胞后会被转录。

Purified DNAs are transcribed after microinjection into Xenopus oocytes.

作者信息

Mertz J E, Gurdon J B

出版信息

Proc Natl Acad Sci U S A. 1977 Apr;74(4):1502-6. doi: 10.1073/pnas.74.4.1502.

Abstract

The possibility of using DNA-injected Xenopus laevis oocytes and eggs for studying the control of transcription in eukaryotes has been investigated. When purified DNA of simian virus 40 (SV40) is injected into Xenopus laevis oocytes, tritiated RNA precursors are incorporated into DNase-I-resistant, RNase-A- and alkali-sensitive material that hybridizes specifically to SV40 DNA. This viral transcription continues for at least 5 days and occurs only when the injected DNA is directed to the nucleus of the oocyte. The quantity of SV40-specific RNA produced is roughly proportional to the amount of DNA injected; above 1 ng per oocyte, most of the nonribosomal RNA made in successfully injected oocytes is virus-specific. Transcription also occurs, although at a lower efficiency, after injection of the DNA into unfertilized eggs. The DNAs of adenovirus 5, cloned Drosophila melanogaster histone genes, and even bacteriophage phiX174 replicative form, bacteriophage phi80plac, and the ColE1 plasmid are also transcribed after injection into oocytes or eggs.

摘要

人们已经研究了利用注射了DNA的非洲爪蟾卵母细胞和卵来研究真核生物转录调控的可能性。当将纯化的猴病毒40(SV40)DNA注射到非洲爪蟾卵母细胞中时,氚标记的RNA前体被掺入到对DNase - I有抗性、对RNase - A和碱敏感的物质中,该物质能与SV40 DNA特异性杂交。这种病毒转录至少持续5天,并且仅当注射的DNA进入卵母细胞核时才会发生。产生的SV40特异性RNA的量大致与注射的DNA量成正比;每个卵母细胞注射超过1 ng时,成功注射的卵母细胞中产生的大多数非核糖体RNA都是病毒特异性的。将DNA注射到未受精卵中后也会发生转录,不过效率较低。腺病毒5的DNA、克隆的果蝇组蛋白基因,甚至噬菌体phiX174复制型、噬菌体phi80plac和ColE1质粒在注射到卵母细胞或卵中后也会被转录。

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本文引用的文献

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The replication of purified DNA introduced into living egg cytoplasm.导入活卵细胞质中的纯化DNA的复制。
Biochim Biophys Acta. 1969 Feb 18;174(2):614-28. doi: 10.1016/0005-2787(69)90291-3.
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Enzymatic end-to end joining of DNA molecules.DNA分子的酶促端对端连接
J Mol Biol. 1973 Aug 15;78(3):453-71. doi: 10.1016/0022-2836(73)90468-3.

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