大鼠血红素加氧酶-1基因在乳酸乳球菌中的表达  

Expression of rat heme oxygenase-1 gene in Lactococcus lactis

作　　者：吴长毅[1] 庞庆丰[2] 曾因明[2] 王俊科[2] 杨光[2] 高新跃[2] 

机构地区：[1]中国医科大学附属第一临床学院麻醉科,辽宁沈阳110001 [2]徐州医院江苏省麻醉学重点实验室,江苏徐州221002

出　　处：《南京医科大学学报（自然科学版）》2006年第2期101-104,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省教育厅开放课题资助项目(KJS04002)

摘　　要：目的:在乳酸乳球菌中表达大鼠血红素加氧酶-1(HO-1)基因。方法:采用RT-PCR技术从大鼠脾总RNA中分离扩增血红素加氧酶HO-1基因,将该基因克隆进pGEM-Teasy质粒中,转化大肠杆菌DH5α提取质粒,鉴定HO-1基因。酶切后与pSEC质粒连接,经电击转化,将重组质粒转入乳酸乳球菌NZ9000中,转化子在含有氯霉素的脑心浸液培养基上培养。用nisin诱导HO-1表达,SDS-PAGE、Westernblot鉴定表达产物,并采用分光光度法测定HO-1活性。结果:表达产物相对分子量约为32kU,表达量约为7.0mg/L,HO-1活性为2.386U/(mg·h)。结论:乳酸乳球菌能够表达具有生物活性的大鼠HO-1。Objective： To clone and express rat heme oxygenase-1 （HO-1） gene in lactococcus lactis. Methods： The rat HO-1 cDNA was amplified by reverse transcription-polymerase chain reaction （RT-PCR）, and the sequence was identified by T/A clone. The HO-1 gene digested by restriction endonuclease was ligated with plasmid pSEC. Then the recombinant plasmid was transformed into lactococcus lactis NZ9000 by electroporation. The expression of heine oxygenase-1 gene induced by nisin was identified by SDS-PAGE and Western blot, and the activity of HO-1 secreted by engineering lactococcus lactis was measured by spectrophotography. Results： The relative molecular weight of expressed protein was about 32 kD, and its expression level was about 7.0 mg/L. The activity of heme oxygenase was 2.386 U/mg per hour. Conclusion： The expression of rat heme oxygenase-1 has been detected in lactococcus lactis.

关 键 词：血红素加氧酶-1 基因克隆表达 乳酸乳球菌 

分 类 号：R344.13[医药卫生—基础医学]