Construction of a human chromosome 3 specific NotI linking library using a novel cloning procedure.
Construction of a human chromosome 3 specific NotI linking library using a novel cloning procedure.
复制标题
使用新型克隆程序构建人类 3 号染色体特异性 NotI 连接文库。
DOI：
10.1093/nar/18.21.6319
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发表时间：
1990
影响因子：
14.9
通讯作者：
Sümegi,J
中科院分区：
文献类型：
--
作者：
Zabarovsky,ER;Boldog,F;Thompson,T;Scanlon,D;Winberg,G;Marcsek,Z;Erlandsson,R;Stanbridge,EJ;Klein,G;Sümegi,J
Two new diphasmid vectors (lambda SK17 and SK22) and a novel procedure to construct linking libraries are described. A partial filling-in reaction provides counter-selection against false linking clones in the library, and obviates the need for supF selection. The diphasmid vectors, in combination with the novel selection procedure, have been used to construct a chromosome 3 specific Notl linking library from a human chromosome 3/mouse microcell hybrid cell line. The application of the new vectors and the strong biochemical and biological selections resulted in a library of 60.000 Notl linking clones. As practically all of them are real Notl linking clones (no false recombinant8) the library represents approximately 3.000 human recombinants (equal to 10–15 genomic equivalents of chromosome 3). Previously published methods for construction of linking libraries are compared with the procedure described in the present paper. The advantages of the new vectors and the novel protocol are discussed.
