Zhou B, Chen N G, Li Q L
Shanghai Institute of Biochemistry, Chinese Academy of Sciences, China.
Gene. 1988 Oct 30;70(2):405-9. doi: 10.1016/0378-1119(88)90213-2.
Partial digestion of a target DNA fragment with 4-bp-recognition restriction enzymes followed by a forced ligation to an M13 vector was employed for the construction of a subfragment library. The library can be used for either shotgun or non-random nucleotide sequencing. Application of the partial digests generated with the 4-bp recognition restriction enzymes instead of DNase I in the improved non-random strategy for nucleotide sequencing (Li and Wu, 1987) made the procedure as easy as that of the random strategy. The library can also be used in shotgun nucleotide sequencing directly, and few self-ligated subfragments were found. The usefulness of this procedure was demonstrated by the sequencing of a goat 6.5-kb EcoRI fragment, which is located 5' to the epsilon globin gene.
用识别4个碱基的限制性内切酶对目标DNA片段进行部分消化，然后强制连接到M13载体上，用于构建亚片段文库。该文库可用于鸟枪法或非随机核苷酸测序。在改进的非随机核苷酸测序策略中（Li和Wu，1987），使用识别4个碱基的限制性内切酶而不是DNase I产生的部分消化产物，使得该过程与随机策略一样简单。该文库也可直接用于鸟枪法核苷酸测序，并且发现很少有自连接的亚片段。通过对位于ε珠蛋白基因5'端的山羊6.5 kb EcoRI片段进行测序，证明了该方法的有效性。
