Heuer T, Bürger C, Maass G, Tümmler B
Klinische Forschergruppe, Zentrum Biochemie, Medizinische Hochschule Hannover, Germany.
Electrophoresis. 1998 Apr;19(4):486-94. doi: 10.1002/elps.1150190406.
Yeast artificial chromosomes (YACs) can accommodate large inserts and hence should be attractive tools for intra- and interspecies comparisons of bacterial genomes. YAC libraries were constructed from size-selected partial digests of human and Pseudomonas aeruginosa PAO DNA and SpeI-restricted PAO DNA. Whereas YACs from human DNA had an average size of 350 kilobase pairs (kbp), a P. aeruginosa sequence larger than 120 kbp was absent or truncated in the eukaryotic host. Coligation occurred for YACs smaller than 40 kbp, but stable YACs with 40-120 kbp large inserts of P. aeruginosa DNA were obtained in high yield. SpeI-restricted chromosomes from 97 P. aeruginosa strains representing 47 genotypes were hybridized with stable YACs from three equidistant regions of the PAO genome. The low complexity of hybridizing bands demonstrated that the analyzed 100 kbp sequence contigs were stably maintained in most P. aeruginosa isolates from both disease and environmental habitats.
酵母人工染色体（YACs）能够容纳大片段插入片段，因此对于细菌基因组的种内和种间比较而言应该是很有吸引力的工具。YAC文库是由人类和铜绿假单胞菌PAO DNA的大小选择部分消化产物以及经SpeI酶切的PAO DNA构建而成。来自人类DNA的YAC平均大小为350千碱基对（kbp），而在真核宿主中，大于120 kbp的铜绿假单胞菌序列缺失或被截断。小于40 kbp的YAC会发生共连接，但能高产获得带有40 - 120 kbp铜绿假单胞菌DNA大片段插入的稳定YAC。来自代表47种基因型的97株铜绿假单胞菌菌株的经SpeI酶切的染色体与来自PAO基因组三个等距区域的稳定YAC进行杂交。杂交带的低复杂性表明，所分析的100 kbp序列重叠群在来自疾病和环境栖息地的大多数铜绿假单胞菌分离株中都能稳定维持。
