Yamaguchi R, Matsuo K, Yamazaki A, Abe C, Nagai S, Terasaka K, Yamada T
Central Research Laboratories, Ajinomoto Co., Inc., Kawasaki, Japan.
Infect Immun. 1989 Jan;57(1):283-8. doi: 10.1128/iai.57.1.283-288.1989.
The gene for immunogenic protein MPB64 found in culture filtrates of only Mycobacterium tuberculosis and some strains of Mycobacterium bovis BCG was cloned by using a single-probe method and was sequenced. The gene analysis revealed that the structural gene for MPB64 consisted of 618 base pairs, and its deduced molecular weight was 22,400. Twenty-two amino acids for a putative signal peptide and 205 amino acids for the MPB64 protein were observed. In the coding region, the third letter of the codon showed a biased codon and a high G+C content (78.5%). The gene was expressed in Escherichia coli by using an E. coli expression vector. The product showed migration similar to that of the authentic MPB64 protein by electrophoresis and reacted with the polyclonal and the monoclonal antibodies raised against the MPB64 protein. The strict specificity of MPB64 could be applied to immunodiagnosis of tuberculosis.
仅在结核分枝杆菌培养滤液以及一些牛型结核分枝杆菌卡介苗菌株中发现的免疫原性蛋白MPB64的基因，采用单探针法进行克隆并测序。基因分析显示，MPB64的结构基因由618个碱基对组成，其推导的分子量为22400。观察到一个推测的信号肽有22个氨基酸，MPB64蛋白有205个氨基酸。在编码区，密码子的第三个字母显示出密码子偏好且G+C含量高（78.5%）。该基因通过使用大肠杆菌表达载体在大肠杆菌中表达。产物经电泳显示出与天然MPB64蛋白相似的迁移情况，并与针对MPB64蛋白产生的多克隆抗体和单克隆抗体发生反应。MPB64的严格特异性可应用于结核病的免疫诊断。
