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用来自米曲霉的核酸酶S1对两种噬菌体λ突变体的DNA异源双链分子进行特异性切割。

Sepcific fragmentation of DNA heteroduplex molecules of two bacteriophage lambda mutants with endonuclease Si from Aspergillus oryzae.

作者信息

Ksënzenko V N, Tikhomirova L P, Matvienko N I

出版信息

Nucleic Acids Res. 1976 Apr;3(4):1081-93. doi: 10.1093/nar/3.4.1081.

Abstract

Heteroduplex DNA molecules of two bacteriophage mutants (lambda b2 and lambda i434ct68) were obtained by the method of molecular hybridization. These heteroduplexes possessed two types of loops formed as a result of: a) deletion in one of the DNA strands; and b) substitution of a DNA fragment for nonhomological one. The digestion of heteroduplexes with single-stranded specific nuclease SI from Aspergillus oryzae produced two fragments at 37 degrees C and three ones at 55 degrees C. The separation of fragments and determination of their molecular weight were carried out by means of electrophoresis in agarose. The molecular weights both measured and preliminarily calculated proved to be close. One of the fragments was identificated by its biological activity in CaCl2-dependent infectious system with helperphage.

摘要

通过分子杂交法获得了两种噬菌体突变体(λb2和λi434ct68)的异源双链DNA分子。这些异源双链体具有两种类型的环,其形成原因如下:a)一条DNA链中的缺失;b)用非同源DNA片段替换。用来自米曲霉的单链特异性核酸酶SI在37℃消化异源双链体产生两个片段,在55℃产生三个片段。通过琼脂糖电泳进行片段的分离及其分子量的测定。测量的分子量和初步计算的分子量证明是接近的。其中一个片段通过其在依赖CaCl2的辅助噬菌体感染系统中的生物学活性进行鉴定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f830/342967/fa20bd87a909/nar00489-0237-a.jpg

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