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本文报道了BCG DNA以λ噬菌体EMBL3为载体在大肠杆菌中的克隆和表达,并描述了大肠杆菌表达的免疫原性蛋白的特征.用限制性核酸内切酶Sau3A将BCG DNA部分切割为10～20千对碱基片段,与Bam HI切割的EMBL3 DNA连接,体外包装后转导至大肠杆菌K-12 LE392株内,结果每μg DNA可产生6000空斑形成单位,使用P2-溶源性大肠杆菌Q359株也可获得同样结果.由于只有重组噬菌体能在P2溶源性细菌中生长,所以试验中获得的空斑是由DNA重组噬菌体所产生.
Here we report the cloning and expression of BCG DNA in lambda phage EMBL3 as a vector in Escherichia coli and characterization of the expressed immunogenic protein in E. coli.The partial BCG DNA was partially cleaved by Sau3A to 10 ~ 20 kb base fragment, ligated to Bam HI-cut EMBL3 DNA, packaged in vitro and transduced into E. coli K-12 LE392 strain. As a result, 6000 plaque-forming units per μg of DNA were produced using P2-lysogenic large intestine The same result was also obtained with the strain of Bacillus strain Q359. Since only recombinant bacteriophages were able to grow in P2 lysogenic bacteria, the plaques obtained in the experiment were produced by recombinant DNA bacteriophages.