摘要
目的利用基因工程方法制备肺炎链球菌噬菌体裂解酶CPL-1。方法根据大肠埃希菌密码子偏爱性合成一段编码cpl-1的基因序列,克隆到表达载体pET28a中,构建出表达质粒pET28a-cpl-1,转化至大肠埃希菌BL21(DE3)中进行表达。结果重组CPL-1在大肠埃希菌中以可溶形式表达,表达量占菌体总蛋白的30%。经胆碱类似物DEAE亲和纯化后,重组CPL-1的纯度大于97%。经体外抑菌活性试验证明其对肺炎链球菌具有明显的抑菌效果。结论噬菌体裂解酶CPL-1的开发利用为预防和治疗肺炎链球菌感染提供了新的方法。
Objective To produce phage lytic enzyme CPL-1 by gene engineering.Methods According to the bias of codon utilization of Escherichia coli,the cpl-1 gene was synthesized and inserted into the vector pET28a to construct the recombinant expression plasmid（pET28a-cpl-1）and then transformed into BL21（DE3）.Results Recombinant CPL-1 was expressed and resulted in soluble form that amounted to 30%（w/w）of total cellular proteins,and then purified by affinity chromatography on DEAE-sepharose.The final recovery of recombinant CPL-1 was 70% and 97% in purity.In vitro susceptibility testing showed that purified CPL-1 had significant antimicrobial activity.Conclusions Development of CPL-1 may provide a new method to prevent and treat S.pneumoniae infection.
出处
《中国感染与化疗杂志》
CAS
2010年第2期139-142,共4页
Chinese Journal of Infection and Chemotherapy
关键词
肺炎链球菌
噬菌体裂解酶
大肠埃希菌
Streptococcus pneumoniae
phage lytic enzyme
Escherichia coli
