cis-trans isomerization of unsaturated fatty acids: Cloning and sequencing of the cti gene from Pseudomonas putida P8
cis-trans isomerization of unsaturated fatty acids: Cloning and sequencing of the cti gene from Pseudomonas putida P8
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DOI：
10.1128/aem.63.11.4292-4297.1997
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发表时间：
1997-11-01
影响因子：
4.4
通讯作者：
Keweloh, H
中科院分区：
文献类型：
--
作者：
Holtwick, R;Meinhardt, F;Keweloh, H
利用mini-Tn 5诱变系统对恶臭假单胞菌P8进行转座子突变，并对突变株进行了耐温性和耐酚性的检测。用5,800株转座子突变株构建了600株温度敏感菌株的库;这些菌株之一被鉴定为其进行脂肪酸顺-反异构化的能力受损，构建了恶臭假单胞菌P8的基因文库，并以其mini-Tn 5插入位点附近的序列为探针进行筛选，分离并克隆了与突变互补的DNA片段。cti基因位于恶臭假单胞菌P8的甲硫氨酸合成酶基因座（metH）附近。将cti基因片段整合到质粒中也赋予大肠杆菌顺反异构化的能力;对cti基因进行了全序列测定，并推导了氨基酸序列。
Transposon mutants of Pseudomonas putida P8 were generated by applying a mini-Tn5 mutagenesis system, The mutants obtained were checked for their ability to tolerate increased temperatures and elevated phenol concentrations, Approximately 5,800 transposon mutants were used to generate a pool of 600 temperature-sensitive strains; one of these strains was identified as being damaged in its ability to perform cis-trans isomerization of fatty acids, A gene library of P. putida P8 was constructed and screened by using as a probe sequences immediately adjacent to the mini-Tn5 insertion, A DNA fragment that complemented the mutation was isolated and cloned. The corresponding gene, termed cti, is located close to the methionine synthase locus (metH) in P, putida P8, A cti-carrying fragment integrated into a plasmid also conferred the ability for cis-trans isomerization to Escherichia coli; the cti gene was completely sequenced, and the amino acid sequence was deduced.