Deckers H M, Voordouw G
Department of Biological Sciences, University of Calgary, Alberta, Canada.
J Bacteriol. 1994 Jan;176(2):351-8. doi: 10.1128/jb.176.2.351-358.1994.
A library of 879 recombinant lambda phages, constructed for the genome of Desulfovibrio vulgaris Hildenborough, has been ordered by restriction fingerprinting. Restriction endonuclease HinfI digestion patterns were entered into a data base and sorted into 87 overlapping groups (contigs), with 19 clones remaining unattached. Eight of ten cloned genes of D. vulgaris, including dcrA, which encodes a transmembrane methyl-accepting protein, were assigned to contigs. Probing of a filter containing the lambda DNAs of the library with the labeled, conserved 3' end of the dcrA gene indicated hybridization to 54 clones distributed over multiple contigs. The presence of 11 additional dcr genes (dcrB to dcrL) was confirmed by direct cycled dideoxy sequencing of positive lambda clones. Since the ordered library provides only partial coverage of the D. vulgaris Hildenborough genome, we estimate that the dcr gene family has 16 members spread throughout the genome, making it the second largest gene family found in prokaryotes.
为普通脱硫弧菌希登伯勒菌株(Desulfovibrio vulgaris Hildenborough)基因组构建的一个包含879个重组λ噬菌体的文库,已通过限制性指纹图谱法进行排序。限制性内切酶HinfI的消化模式被录入数据库,并被分类为87个重叠组(重叠群),有19个克隆未被归入其中。普通脱硫弧菌的10个克隆基因中有8个,包括编码跨膜甲基接受蛋白的dcrA,被归入重叠群。用dcrA基因标记的保守3'末端对包含文库λ DNA的滤膜进行杂交检测,结果显示与分布在多个重叠群中的54个克隆发生杂交。通过对阳性λ克隆进行直接循环双脱氧测序,证实了另外11个dcr基因(dcrB至dcrL)的存在。由于该排序文库仅提供了普通脱硫弧菌希登伯勒菌株基因组的部分覆盖,我们估计dcr基因家族有16个成员分布在整个基因组中,使其成为在原核生物中发现的第二大基因家族。