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1980年美国遗传学会会议论文摘要。科罗拉多州博尔德,1980年8月18日至20日。

Abstracts of papers presented at the 1980 meetings of the Genetic Society of America. Boulder, Colorado August 18-20, 1980.

作者信息

Chumley Forrest G., Roth John R.

机构信息

Department of Biology, University of Utah, Salt Lake City, Utah 84112

出版信息

Genetics. 1980;94(4 Pt 2 Suppl):1-16.

Abstract

The transposable tetracycline resistance element, Tn10, can serve as a region of homology to promote rec-dependent deletion, duplication and directed transposition of bacterial genes. Tn10 insertions in regions of the chromosome near the histidine operon (his) have been isolated and characterized in Salmonella typhimurium. When strains are constructed containing two Tn10 insertions flanking the his operon in the same orientation (Tn10-his-Tn10), recombination can occur between Tn10 sequences resulting in the deletion of the intervening his region. The sites of the Tn10 insertions determine the endpoints of the deletion. In crosses designed to construct strains carrying Tn10-his-Tn10, another class of unstable recombinants arises in which the his region exists in tandem duplication, with a Tn10 insertion joining the duplicated copies (his-Tn10-his). The sites of the parental Tn10 insertions mark the endpoints of the duplication. When a strain carrying Tn10-his-Tn10 is used as a donor of his(+) in conjugation or P22-mediated transduction, recombinants can arise in which the his region has been transposed to the site of any Tn10 insertion, far from the normal location of his in the recipient chromosome. In this manner, the his operon has been moved to the site of a pyrB::Tn10 insertion and has been placed on F' plasmids. At these new locations, the his(+) character shows the rec-dependent deletion of his(+) expected for a Tn10-his-Tn10 duplication. These methods should be generally useful for the manipulation of bacterial genes.

摘要

可转座的四环素抗性元件Tn10可作为同源区域,促进细菌基因的rec依赖性缺失、重复和定向转座。已在鼠伤寒沙门氏菌中分离并鉴定了组氨酸操纵子(his)附近染色体区域的Tn10插入。构建在his操纵子两侧以相同方向含有两个Tn10插入的菌株(Tn10-his-Tn10)时,Tn10序列之间可发生重组,导致中间的his区域缺失。Tn10插入位点决定了缺失的端点。在设计构建携带Tn10-his-Tn10菌株的杂交实验中,出现了另一类不稳定重组体,其中his区域以串联重复形式存在,一个Tn10插入连接了重复拷贝(his-Tn10-his)。亲本Tn10插入位点标记了重复的端点。当携带Tn10-his-Tn10的菌株在接合或P22介导的转导中用作his(+)供体时,可产生重组体,其中his区域已转座到任何Tn10插入位点,远离其在受体染色体中的正常位置。通过这种方式,his操纵子已被转移到pyrB::Tn10插入位点,并被置于F'质粒上。在这些新位置,his(+)性状表现出与Tn10-his-Tn10重复预期的rec依赖性his(+)缺失。这些方法通常应有助于细菌基因的操作。

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