农业科学与技术（英文版）
農業科學與技術（英文版）
농업과학여기술（영문판）
AGRICULTURAL SCIENCE & TECHNOLOGY
2010年
5期
55-58,86
,共5页
李飞武%邵改革%邢珍娟%李葱葱%夏蔚%张明
李飛武%邵改革%邢珍娟%李蔥蔥%夏蔚%張明
리비무%소개혁%형진연%리총총%하위%장명
转基因生物%质粒标准分子%MON89788大豆%转化体特异性检测
轉基因生物%質粒標準分子%MON89788大豆%轉化體特異性檢測
전기인생물%질립표준분자%MON89788대두%전화체특이성검측
Genetically modified organisms%Plasmid reference molecule%MON89788 soybean%Event-specific detection
[目的]构建适用于转基因大豆MON89788检测的质粒标准分子.[方法]利用定性PCR和连接、转化等分子克隆技术,将大豆内标准基因lectin、MON89788的3'端特异性序列和5'端特异性序列依次克隆到pMD18-T载体上,获得质粒标准分子pMD-LM3M5,并进行适用性验证.[结果]获得了3 700 bp的质粒标准分子,其中重组DNA片段1 029 bp.该质粒标准分子的定性PCR检测灵敏度达到10 copy.[结论]该研究构建的质粒标准分子pMD-LM3M5能替代NON89788基体标准品,用于MON89788大豆及其产品的定性PCR检测.
[目的]構建適用于轉基因大豆MON89788檢測的質粒標準分子.[方法]利用定性PCR和連接、轉化等分子剋隆技術,將大豆內標準基因lectin、MON89788的3'耑特異性序列和5'耑特異性序列依次剋隆到pMD18-T載體上,穫得質粒標準分子pMD-LM3M5,併進行適用性驗證.[結果]穫得瞭3 700 bp的質粒標準分子,其中重組DNA片段1 029 bp.該質粒標準分子的定性PCR檢測靈敏度達到10 copy.[結論]該研究構建的質粒標準分子pMD-LM3M5能替代NON89788基體標準品,用于MON89788大豆及其產品的定性PCR檢測.
[목적]구건괄용우전기인대두MON89788검측적질립표준분자.[방법]이용정성PCR화련접、전화등분자극륭기술,장대두내표준기인lectin、MON89788적3'단특이성서렬화5'단특이성서렬의차극륭도pMD18-T재체상,획득질립표준분자pMD-LM3M5,병진행괄용성험증.[결과]획득료3 700 bp적질립표준분자,기중중조DNA편단1 029 bp.해질립표준분자적정성PCR검측령민도체도10 copy.[결론]해연구구건적질립표준분자pMD-LM3M5능체대NON89788기체표준품,용우MON89788대두급기산품적정성PCR검측.
[Objective] The aim was to construct a plasmid reference molecule (PRM) for detection of transgenic soybean MON89788.[Method]the lectin gene sequence,3'-junction and 5'-junction sequence between host plant DNA integrated DNA of MON89788 soybean were amplified independently,and the three fragments were cloned into the cloning vector pMD18-T in order through molecular manipulation method to construct pMD-LM3M5,the applicability of the constructed novel PRM was tested. [Result] Sequencing confirmation result showed that the PRM was 3 700 bp in length,containing 1 029 bp of recombined DNA fragment.The limits of qualitative detection of the PRM were 10 copies,[Conclusion]The PRM constructed in this study was suitable for the identification of MON89788 event.
