CHLORAMPHENICOL RESISTANCE IN CAMPYLOBACTER-COLI - NUCLEOTIDE-SEQUENCE, EXPRESSION, AND CLONING VECTOR CONSTRUCTION
CHLORAMPHENICOL RESISTANCE IN CAMPYLOBACTER-COLI - NUCLEOTIDE-SEQUENCE, EXPRESSION, AND CLONING VECTOR CONSTRUCTION
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DOI：
10.1016/0378-1119(90)90463-2
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发表时间：
1990-09-28
期刊：
影响因子：
3.5
通讯作者：
TAYLOR, DE
中科院分区：
文献类型：
--
作者：
WANG, Y;TAYLOR, DE
从日本大肠弯曲杆菌pNR9589质粒中分离得到一株氯霉素耐药决定因子（CmR），并测定其核苷酸序列，全长621 bp。该基因产物被鉴定为Cm乙酰转移酶（CAT），其氨基酸序列与其他革兰氏+和革兰氏-来源的CAT蛋白有43% ~ 57%的同源性。虽然在大肠杆菌和大肠杆菌中cat基因的表达是组成性的，但引物延伸实验的结果表明，在这两个物种中，转录是在不同的位点开始的。一个卡那霉素耐药的决定因素，被鉴定为aphA-3基因，位于cat基因的下游。cat基因的用途与大肠杆菌的用途大不相同，但在大肠杆菌的大细胞中大量合成了cat多肽。因此，密码子使用偏差不是影响弯曲杆菌基因在大肠杆菌中表达的障碍之一。本研究构建了新的弯曲杆菌克隆载体。
A chloramphenicol-resistance determinant (CmR), originally cloned from Campylobacter coli plasmid pNR9589 in Japan, was isolated and the nucleotide sequence determined, which contained an open reading frame of 621 bp. The gene product was identified as Cm acetyltransferase (CAT), which had a putative amino acid sequence that showed 43% to 57% identity with other CAT proteins of both Gram+ and Gram- origin. Although expression of the cat gene was constitutive in both C. coli and Escherichia coli, results of primer extension experiments indicated that transcription was initiated at different sites in these two species. A kanamycin-resistance determinant, identified as the aphA-3 gene, was located downstream from the cat gene. The usage of the cat gene is very different from that used in E. coli, however, the CAT polypeptide was synthesized in large amounts in E. coli maxicells. Therefore, the codon usage bias is not one of the obstacles which affects Campylobacter spp. gene expression in E. coli. New Campylobacter cloning vectors were constructed in this study.