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A phagemid vector library for cloning DNA with four-nucleotide 5' or 3' overhangs.

Abstract：

:A phagemid vector library for cloning DNA with four nucleotide 5' or 3' overhangs has been constructed. This library is based on the pT7T3 vector (Pharmacia) which is a modification of the phagemid pTZ18U vector. We have chosen pT7T3 as the parent vector because it can be used for Sanger's dideoxy sequencing and for the generation of RNA probes with either the T7 or T3 promoter. Each member of the cloning vector series pBM has recognition sites for both of the restriction enzymes BspM1 and BstX1 in addition to the basic multiple cloning sites. BspM1 recognizes the sequence 5'...ACCTGC NNNN/NNNN...3' whereas BstX1 recognizes the sequence 5'...CCAN NNNN/NTGG...3'. Thus these two sites can be overlapped, so that only 256 vectors (instead of 512 vectors) need be constructed to cover all the theoretical possible combinations of sites which give complementary cohesive ends for cloning DNA with four nucleotide 5' or 3' overhangs. This vector library can be used for amplification cloning of DNA in a tandem array by choosing appropriate vectors which have nonpalindromic sequences. We have obtained approximately 200 members of the 256 possible clones and have organized the vectors using a MacIntosh HyperCard program for easy retrieval.

journal_name

Plasmid

journal_title

Plasmid

authors

Waye MM,Mui F,Hodge K,Li VK

doi

10.1016/0147-619x(91)90038-x

subject

Has Abstract

pub_date

1991-07-01 00:00:00

pages

74-7

issue

1

eissn

0147-619X

issn

1095-9890

pii

0147-619X(91)90038-X

journal_volume

26

pub_type

杂志文章

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