山东医药
山東醫藥
산동의약
SHANDONG MEDICAL JOURNAL
2015年
4期
4-7
,共4页
郭永红%问娇%王俊红
郭永紅%問嬌%王俊紅
곽영홍%문교%왕준홍
Adnectin%PCR扩增%基因合成%核糖体展示文库
Adnectin%PCR擴增%基因閤成%覈糖體展示文庫
Adnectin%PCR확증%기인합성%핵당체전시문고
Adnectin%PCR amplification%gene synthesis%ribosomes display library
目的：构建Adnectin半随机的核糖体（ mRNA）展示文库。方法分析Adnectin核糖体展示文库结构序列的编码氨基酸序列，利用无意突变建立酶切位点，使用PCR扩增和基因合成两种方法相结合，构建Adnectin半随机核糖体展示文库，通过限制性内切酶及DNA测序证实序列正确性，对文库转化菌的滴定和插入失活蓝白菌斑筛查和计数测定计算文库的库容。结果经测序证实文库结构的正确性，文库的库容为1．54×1013／mL。结论成功构建Adnectin半随机的核糖体展示文库，方法简单，库容量大，该文库的建成为亲和各种相关蛋白的Adnectin结合序列奠定基础。
目的：構建Adnectin半隨機的覈糖體（ mRNA）展示文庫。方法分析Adnectin覈糖體展示文庫結構序列的編碼氨基痠序列，利用無意突變建立酶切位點，使用PCR擴增和基因閤成兩種方法相結閤，構建Adnectin半隨機覈糖體展示文庫，通過限製性內切酶及DNA測序證實序列正確性，對文庫轉化菌的滴定和插入失活藍白菌斑篩查和計數測定計算文庫的庫容。結果經測序證實文庫結構的正確性，文庫的庫容為1．54×1013／mL。結論成功構建Adnectin半隨機的覈糖體展示文庫，方法簡單，庫容量大，該文庫的建成為親和各種相關蛋白的Adnectin結閤序列奠定基礎。
목적：구건Adnectin반수궤적핵당체（ mRNA）전시문고。방법분석Adnectin핵당체전시문고결구서렬적편마안기산서렬，이용무의돌변건립매절위점，사용PCR확증화기인합성량충방법상결합，구건Adnectin반수궤핵당체전시문고，통과한제성내절매급DNA측서증실서렬정학성，대문고전화균적적정화삽입실활람백균반사사화계수측정계산문고적고용。결과경측서증실문고결구적정학성，문고적고용위1．54×1013／mL。결론성공구건Adnectin반수궤적핵당체전시문고，방법간단，고용량대，해문고적건성위친화각충상관단백적Adnectin결합서렬전정기출。
Objective To construct semi-random Adnectin ribosome display library .Methods Adnectin ribosome display library was constructed by PCR amplification combining with gene synthesis method .PCR products contained the restrictive sites of SpeⅠbased on nonsense mutation .The recombinant plasmids were identified by restriction endonuclease enzyme analysis and DNA sequence .The positive recombinant clones were screened and library capacity was measured by blue-white test.Result The library was confirmed by sequencing and library capacity was 1.54 ×1013/mL.Conclusions To corstruct successful Adnectin semi-random ribosomes display library ( mRNA) used simple and easy method .The built library laid the foundation for Adnectin affinity with various related protein .
