The complete amino acid sequence of the Clostridium botulinum type D neurotoxin, deduced by nucleotide sequence analysis of the encoding phage d-16 phi genome.
The complete amino acid sequence of the Clostridium botulinum type D neurotoxin, deduced by nucleotide sequence analysis of the encoding phage d-16 phi genome.
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D 型肉毒梭菌神经毒素的完整氨基酸序列，通过编码噬菌体 d-16 phi 基因组的核苷酸序列分析推导出来。
DOI：
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发表时间：
1992
影响因子：
1.2
通讯作者：
K. Inoue
中科院分区：
文献类型：
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作者：
H. Sunagawa;T. Ohyama;T. Watanabe;K. Inoue
测定了D型肉毒梭菌CB16神经毒素的完整核苷酸序列，并首次报道了推导的氨基酸序列。从分子生物学的角度讨论了D型肉毒杆菌神经毒素的结构和功能。从 D 型肉毒杆菌 CB16 菌株的毒素转化噬菌体 d-16 phi 中提取 DNA，并使用 lambda gt11 将编码神经毒素的片段（约 10 kbp）克隆到大肠杆菌中。合成了对应于D型神经毒素N末端附近部分氨基酸序列的Phe7至Val13的21聚体寡核苷酸，并用作探针来鉴定编码D型神经毒素的基因。该核苷酸序列包含编码1,275个氨基酸（分子量为146,785）的单个开放阅读框，推导的氨基酸序列与通过神经毒素片段直接微测序确定的部分氨基酸序列完全一致。在神经毒素的双链分子中，Thr2和Asn443分别形成轻链（分子量50,410）和重链（分子量96,394）的N端，这两条链通过轻链上的Cys437和重链上的Cys450之间的二硫键连接。 D-CB16 神经毒素的核苷酸序列与之前报道的 D 型神经毒素的核苷酸序列有 3 个核苷酸不同。
The complete nucleotide sequence of Clostridium botulinum type D strain CB16 neurotoxin was determined and the deduced amino acid sequence is reported here for the first time. The structure and function of botulinum type D neurotoxin is discussed from a molecular biological viewpoint. DNA was extracted from toxin-converting phage d-16 phi of C. botulinum type D strain CB16, and a fragment (about 10 kbp) coding for the neurotoxin was cloned into Escherichia coli using lambda gt11. A 21-mer oligonucleotide which corresponds to Phe7 to Val13 of the partial amino acid sequence near the N-terminus of the type D neurotoxin was synthesized and used as a probe to identify the gene encoding type D neurotoxin. The nucleotide sequence contained a single open reading frame coding for 1,275 amino acids (molecular weight of 146,785) and the deduced amino acid sequence corresponded exactly to the partial amino acid sequences determined by direct microsequencing of the neurotoxin fragments. In the dichain molecule of the neurotoxin, Thr2 and Asn443 formed the N-termini of the light chain (M.W. 50,410) and heavy chain (M.W. 96,394) respectively, and these two chains were linked with a disulfide bond between Cys437 on the light chain and Cys450 on the heavy chain. The nucleotide sequence of the D-CB16 neurotoxin differed from that previously reported for type D neurotoxin by three nucleotides.