Cao C, Shi C, Li P, Ma Q
Institute of Biotechnology, Academy of Military Medical Sciences, Beijing.
Yi Chuan Xue Bao. 1997;24(1):78-86.
A promoter sequence, which promotes the transcription of cholera toxin B subunit gene, was found in cholera toxin A subunit structure gene. The transcription starts at the adenine Located at +833, that is 456bp upstream to the A of the initiation codon ATG of cholera toxin B gene. Under the control of the promoter, cholera toxin B subunit was over-expressed as high as 200 mg/L at an optimized culture condition. The chloramphenicol acetyl transferase gene and beta-galactosidase could also be efficiently expressed under the direction of the promoter. This promoter may be responsible for the 6 fold and 7 fold higher expression level of cholera toxin B subunit than cholera toxin A subunit in V. cholerae and Escheria coli respectively. The over-expression of CTB may be useful in preparing vaccine against cholera and facilitating the construction of peptide-bearing immunogenic hybrid proteins.
在霍乱毒素A亚基结构基因中发现了一个可促进霍乱毒素B亚基基因转录的启动子序列。转录起始于位于+833的腺嘌呤,该位置在霍乱毒素B基因起始密码子ATG的A上游456bp处。在该启动子的控制下,在优化的培养条件下,霍乱毒素B亚基的表达量高达200mg/L。氯霉素乙酰转移酶基因和β-半乳糖苷酶在该启动子的指导下也能高效表达。该启动子可能分别是霍乱毒素B亚基在霍乱弧菌和大肠杆菌中的表达水平比霍乱毒素A亚基高6倍和7倍的原因。霍乱毒素B亚基的过表达可能有助于制备霍乱疫苗,并促进含肽免疫原性杂合蛋白的构建。