Multi-primer Multiplex PCR Detection Method for the Exogenous Multivalent Insect-resistant Bt Genes in Transgenic Rice

doi:10.13560/j.cnki.biotech.bull.1985.2019-0383

Abstract

Abstract: This work aims to explore a simple and effective method for detecting multivalent insect-resistant Bt genes in transgenic rice. Based on the insertion positions of 3 Bt genes（cry1Ab/Ac,cry2,and cry1C）from events（TT51-1,T2A-1,and T1C-19）,9 primers were designed and used to obtain 6 target bands in significant differences by multiplex primer PCR technique. Subsequently,the positive and negative bands representing the TT51-1 event were 937 bp and 718 bp,those representing the T2A-1 event were 600 bp and 434 bp,and those representing the T1C-19 event were 495 bp and 792 bp. The detection of homozygous,heterozygous and negative Bt transgenic rice plants was rapidly conducted by agarose gel electrophoresis. This technology overcame the technical difficulties such as mutual binding interference among the multiple primers in the PCR process and presented fine compatibility with different PCR reagents. It can cooperate with rapid DNA extraction of rice to rapidly analyze a large number of breeding offspring. In conclusion,the establishment of this method provides a simple and efficient gene detection technology for rice stem borer-resistant molecular breeding with multivalent Bt gene,which is beneficial to increase the detection efficiency of Btgenes and to accelerate the breeding process of rice stem borer-resistant transgenic rice.

Key words: Bt gene, transgenic rice, multi-primer multiplex PCR, molecular detection

FANG Xin-yi, YANG Jing, WANG Jing-zhang, LI Yang-sheng. Multi-primer Multiplex PCR Detection Method for the Exogenous Multivalent Insect-resistant Bt Genes in Transgenic Rice[J]. Biotechnology Bulletin, 2019, 35(12): 175-183.

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