Alteration of open reading frames by use of new gene cassettes.
Alteration of open reading frames by use of new gene cassettes.
复制标题
通过使用新的基因盒改变开放阅读框。
DOI：
10.1006/abio.1999.4004
复制
发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Phillips,GJ
中科院分区：
文献类型：
--
作者：
Phillips,GJ
A number of plasmids that carry antibiotic resistance (AbR) 1 genes flanked by restriction sites (gene cassettes) have been described (1–17). Although the primary use of these AbR cassettes has been for genetic analysis of bacteria by insertional disruption of genes in vitro, this report describes the construction and use of new AbR gene cassettes as a means to facilitate construction of gene fusions by altering the reading frame of specific coding regions. One limitation of many of the available gene cassettes is that they do not always represent a variety of AbR markers, nor do they consistently offer a wide selection of restriction sites that flank the resistance gene. Furthermore, many of the available AbR genes also have naturally occurring restriction sites within the coding region, making them unusable for cloning an intact resistance gene. We have previously reported a series of plasmids that overcome many of these limitations in that they carry different AbR cassettes that are symmetrically flanked by a variety of unique restriction sites (13). These cassettes still, however, retain a limited order of restriction enzyme recognition sites with respect to the AbR marker. An AbR cassette excised by most enzymes will also carry with it any number of additional restriction sites that are internal to the site used for excision. These additional restriction sites are, consequently, introduced to a gene construct along with the AbR marker. These additional restriction sites may be undesirable in that they can limit the choice of restriction enzymes that can be used for subsequent cloning steps.
