| pQE31-HPV16 L1重组质粒的构建及鉴定 |
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| 引用本文： | 商庆龙,隋丽华,李迪,王晶,钟照华,谷鸿喜.pQE31-HPV16 L1重组质粒的构建及鉴定[J].哈尔滨医科大学学报,2001,35(1):7-9. |
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| 作者姓名： | 商庆龙 隋丽华 李迪 王晶 钟照华 谷鸿喜 |
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| 作者单位： | 1. 哈尔滨医科大学 微生物学教研室，黑龙江 哈尔滨 150086
2. 哈尔滨医科大学附属第三医院 妇科，黑龙江 哈尔滨 150040 |
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| 基金项目： | 黑龙江省科委攻关课题!(G98 C0 80 3 ) |
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| 摘 要： | 目的 构建重组原核表达质粒以获得ＨＰＶ１６ Ｌ１活性蛋白。为进一步研制ＨＰＶ１６基因工程疫苗打下基础。方法 以克隆质粒pQE３１－ＨＰＶ１６Ｌ１重组质粒，用酶切电泳验证重组结果的正确性，测序检查质闰重组后序列情况。结果 ＰＣＲ结果显示扩增片断大小为１.5Kb,与预期相同。重组质粒酶切后显示其大小约５.0Kb。大小及酶切图谱与预期相同。经测序发现插入片段两端序列无改变。结论 pQE３１－ＨＰＶ１６Ｌ１质粒构建成功。
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| 关 键 词： | 人乳头瘤病毒16型 原核表达系统 pQE31－HPV16L1 |
| 文章编号： | 1000-1905(2001)01-0007-03 |
| 修稿时间： | 2000年10月16 |
Construction and identification of recombinant plasmid pQE31-HPV16 L1 |
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| SHANG Qing long ,SUI Li hua ,LI Di ,et al.Construction and identification of recombinant plasmid pQE31-HPV16 L1[J].Journal of Harbin Medical University,2001,35(1):7-9. |
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| Authors: | SHANG Qing long SUI Li hua LI Di |
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| Institution: | SHANG Qing long 1,SUI Li hua 2,LI Di 1,et al |
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| Abstract: | Objective To construct a recombinant plasmid pQE31 HPV16 L1 for obtaining the HPV16 L1 protein expressed in E.coli and developing the genetically engineered vaccine.Methods PCR was used to amplify the HPV16 L1 gene from plasmid pCR2.1 HPV16 L1 in which the HPV16 L1 was cloned. A new plasmid, pQE31 HPV16 L1, was then constructed by inserting the amplified HPV16 L1 gene into pQE31, a prokaryotic expression vector. Restriction analysis and sequencing were used to confirm the structure of pQE31 HPV16 L1.Results A DNA fragment in size of 1.5Kb was amplified. Restriction analysis showed that the amplified gene was inserted into pQE31 correctly. Sequencing showed there was no mutation in both ends of the inserted L1 gene.Conclusion The plasmid pQE31 HPV16 L1 is constructed successfully. |
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| Keywords: | human papillomavirus type 16 L1 pQE31 prokaryotic expression system |
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