Campbell K, Collins M D, East A K
Department of Microbiology, Institute of Food Research, Reading Laboratory, Earley Gate, UK.
Biochim Biophys Acta. 1993 Dec 14;1216(3):487-91. doi: 10.1016/0167-4781(93)90020-e.
The neurotoxin gene from Clostridium botulinum type G was cloned as a series of overlapping DNA fragments generated using polymerase chain reaction (PCR) technology and primers designed to conserved regions of published botulinal toxin (BoNT) sequences. The 5'-end of the gene was obtained using a primer based on a conserved region of the nontoxic-nonhaemagglutinin gene lying upstream of the toxin gene. Translation of the nucleotide sequence derived from the cloned PCR fragments demonstrated that the gene encodes a protein of 1297 amino acid residues (rmm 149, 147). Comparative alignment of the determined BoNT/G sequence with those of other clostridial neurotoxins revealed highest sequence relatedness (approx. 58% amino acid identity) with BoNT/B of proteolytic and non-proteolytic C. botulinum. Tetanus toxin (TeTx) and other BoNT types revealed lower levels of relatedness with BoNT/G (approximate range 35-42% amino acid identity).
利用聚合酶链反应(PCR)技术和针对已发表肉毒杆菌毒素(BoNT)序列保守区域设计的引物,将来自G型肉毒梭菌的神经毒素基因克隆为一系列重叠的DNA片段。该基因的5'端是使用基于毒素基因上游无毒非血凝素基因保守区域的引物获得的。对克隆的PCR片段推导的核苷酸序列进行翻译表明,该基因编码一个由1297个氨基酸残基组成的蛋白质(相对分子质量149,147)。将确定的BoNT/G序列与其他梭菌神经毒素的序列进行比较比对,结果显示与蛋白水解性和非蛋白水解性肉毒梭菌的BoNT/B具有最高的序列相关性(约58%的氨基酸同一性)。破伤风毒素(TeTx)和其他BoNT类型与BoNT/G的相关性较低(氨基酸同一性约为35-42%)。