ISOLATION AND DIRECT COMPLETE NUCLEOTIDE DETERMINATION OF ENTIRE GENES - CHARACTERIZATION OF A GENE CODING FOR 16S-RIBOSOMAL RNA
ISOLATION AND DIRECT COMPLETE NUCLEOTIDE DETERMINATION OF ENTIRE GENES - CHARACTERIZATION OF A GENE CODING FOR 16S-RIBOSOMAL RNA
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DOI：
10.1093/nar/17.19.7843
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发表时间：
1989-10-11
影响因子：
14.9
通讯作者：
BOTTGER, EC
中科院分区：
文献类型：
--
作者：
EDWARDS, U;ROGALL, T;BOTTGER, EC
使用一组与广泛保守序列同源的合成寡核苷酸，通过聚合酶链反应进行体外扩增，然后直接测序，几乎可以确定编码16S核糖体RNA的基因的完整核苷酸。作为模型系统，测定了堪萨斯分枝杆菌16S rRNA基因的核苷酸序列，发现与牛分枝杆菌BCG同源性为98.7%。这是首次在没有任何亚克隆程序的情况下获得全长1.5 kb的完整扩增基因的连续序列信息。
Using a set of synthetic oligonucleotides homologous to broadly conserved sequences in-vitro amplification via the polymerase chain reaction followed by direct sequencing results in almost complete nucleotide determination of a gene coding for 16S ribosomal RNA. As a model system the nucleotide sequence of the 16S rRNA gene of M. kansasii was determined and found to be 98.7% homologous to that of M. bovis BCG. This is the first report on a contiguous sequence information of an entire amplified gene spanning 1.5 kb without any subcloning procedures.