| 卡介苗穿梭表达质粒pMSIL-2的构建及鉴定 |
| |
| 引用本文： | 刘海涛,孙晓文,李博,鲁军,朱江,凡杰,夏术阶.卡介苗穿梭表达质粒pMSIL-2的构建及鉴定[J].上海医学,2004,27(5):309-311. |
| |
| 作者姓名： | 刘海涛 孙晓文 李博 鲁军 朱江 凡杰 夏术阶 |
| |
| 作者单位： | 200080,上海交通大学附属上海市第一人民医院泌尿外科 |
| |
| 基金项目： | 上海市科委重大项目研究资助 ( 0 3 4119849) |
| |
| 摘 要： | 目的 构建分泌性表达白介素(IL)-2的重组卡介苗(BCG)。方法 分别以BCG和IL-2cDNA为模板，通过PCR扩增得到约117bp的BCG抗原85B(BCG-Ag85B)信号肽序列和399bp的IL-2基因序列。将BCG-Ag85B信号肽序列克隆至大肠杆菌-BCG穿梭质粒pMV261，得到重组质粒pMS。再将IL-2基因克隆至pMS中，得到重组质粒pMSIL-2。结果 质粒pMSIL-2经双酶切和PCR扩增及测序鉴定证实，克隆基因BCG-Ag85B信号肽和IL-2正确插入载体pMV261。结论 重组质粒pMSIL-2可望在BCG中分泌性表达细胞因子IL-2，该质粒的构建成功为改造BCG、发展新型抗膀胱肿瘤疫苗奠定了基础。
|
| 关 键 词： | 白介素-2 重组穿梭质粒 PCR扩增 卡介苗 |
Construction and identification of recombinant shuttle-plasmid with secretory interleukin-2 |
| |
| LIU Haitao,SUN Xiaowen,LI Bo,et al..Construction and identification of recombinant shuttle-plasmid with secretory interleukin-2[J].Shanghai Medical Journal,2004,27(5):309-311. |
| |
| Authors: | LIU Haitao SUN Xiaowen LI Bo |
| |
| Institution: | LIU Haitao,SUN Xiaowen,LI Bo,et al. Department of Urology,Shanghai First People's Hospital,Jiaotong University,Shanghai 200080,China |
| |
| Abstract: | Objective To construct a recombinant secretory BCG Ag85B fused human interleukin (IL) 2. Methods BCG Ag85B signal sequence and IL 2 gene were amplified from the genome of BCG and IL 2 by PCR, respectively. BCG Ag85B signal sequence was cloned in E.coli BCG shuttle vector pMV261 to obtain pMS. Then a new recombinant plasmid pMSIL 2 was constructed by inserting the IL 2 gene into pMS. Results The cloned genes BCG Ag85B and IL 2 were correctly inserted into the vector pMV261,which had been confirmed by restriction endonuclease digestion and PCR amplification of IL 2 and gene sequencing, respectively. Conclusion pMSIL 2 is expected to possess expression of secretory IL 2 in BCG providing the basis for the development of a new anti bladder cancer vaccine. |
| |
| Keywords: | BCG Interleukin 2 Recombinant shuttle plasmid PCR amplification |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
