利用寡脱氧核苷酸定向诱变构建改良的M13载体。

Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis.

作者信息

Norrander J, Kempe T, Messing J

出版信息

Gene. 1983 Dec;26(1):101-6. doi: 10.1016/0378-1119(83)90040-9.

Abstract

The restriction endonuclease cleavage sites for SphI and KpnI have been added to the lac cloning region of the phage vectors M13mp10 and M13mp11, using oligodeoxynucleotide-directed in vitro mutagenesis. Complementary deoxy 16-, 21- or 18-mers with the desired base changes were annealed to the M13mp DNA strand and extended with the Klenow fragment of DNA polymerase I. In adding these sites we have shown that this technique can be used as a general method for inserting sequences of DNA as well as introducing deletions and base pair changes.

摘要

利用寡聚脱氧核苷酸定向体外诱变技术，已将SphI和KpnI的限制性内切酶切割位点添加到噬菌体载体M13mp10和M13mp11的lac克隆区域。将具有所需碱基变化的互补16聚体、21聚体或18聚体脱氧核苷酸与M13mp DNA链退火，并使用DNA聚合酶I的Klenow片段进行延伸。在添加这些位点的过程中，我们已经证明该技术可用作插入DNA序列以及引入缺失和碱基对变化的通用方法。