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[使用黏粒构建集胞藻6803基因组文库]

[Construction of the genomic library for cyanobacteria Synechocystis sp. PCC 6803 using a cosmid].

作者信息

Shestopalov V I, Nashchokina O O, Shestakov S V, Iankovskiĭ N K

出版信息

Genetika. 1994 Apr;30(4):452-5.

PMID:8045394
Abstract

The gene library of the phototrophic cyanobacterium Synechocystis PCC6803 was constructed in the cosmid vector Lorist6. About -3 x 10(3) recombinant clones were obtained, of which 760 were selected for further work. Each clone was kept in an individual well of a microtiter plate. The clone grids obtained were hybridized with two DNA probes, which contained the ndhE and dfr genes of Synechocystis PCC6803. Two groups of probe-overlapping cosmids (11 and 8 clones respectively), containing insertions averaging of 39 kb, were isolated. The capacity of the gene library is nine equivalents of the Synechocystis PCC6803 genome and its representativity exceeds 99%.

摘要

光合蓝细菌聚球藻PCC6803的基因文库构建于黏粒载体Lorist6中。获得了约3×10³个重组克隆,从中挑选出760个用于进一步研究。每个克隆保存在微量滴定板的一个单独孔中。将得到的克隆网格与两个DNA探针杂交,这两个探针包含聚球藻PCC6803的ndhE和dfr基因。分离出两组探针重叠的黏粒(分别为11个和8个克隆),其插入片段平均长度为39 kb。该基因文库的容量为聚球藻PCC6803基因组的9倍,其代表性超过99%。

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