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Food Control
Vol. 62, 2016, Pages: 357–364

A rapid loop-mediated isothermal amplification method for detection of the modified GM cry1A gene in transgenic insect-resistant cotton and rice

Peili Shen, Fengzhen Geng, Yan Yu, Yunzhe Zhang, Zhixin Wang, Zhihui Li, Wei Zhang, Changlong Shu, Yongjun Zhang, Jianxin Tan

College of Food Science, Agricultural Products Processing Engineering Technology Center of Hebei, Agricultural University of Hebei, Baoding 071001, PR China.

Abstract

Among the commercial genetically modified (GM) crops, the insect-resistant GM crops are the major cultivars that cry gene is introduced into. A cry1Ab/1Ac GM fusion gene (GFM cry1A) and a GM truncated cry1Ac gene (cry1Ac-Mon) is the key foreign gene employed for construction of GM crops by China researchers and Monsanto Technology LLC respectively. Here these two genes are entitled “GM cry1A” gene and a rapid visual loop-mediated isothermal amplification (LAMP) assay method for detection of GM cry1Ain transgenic insect-resistant crops was established. The LAMP assay was performed at an optimal temperature of 65 °C for 60 min in the presence of a set of four specific primers recognized six distinct sequences of the GM cry1A gene. The rough detection limit to the GM cry1A in samples is as low as 0.01% (a weight ratio of transgenic insect-resistant rice/cotton to non-transgenic rice/cotton). Comparatively, the sensitivity of this LAMP method is 10 times over that of the conventional PCR method. Fifteen cultivars/events and five Bt strains with or without cry1A gene were analyzed using the LAMP method as well as PCR method. The results demonstrate that this LAMP method shows a distinct specificity to the GM cry1A gene comparing with PCR analysis. Therefore, this LAMP method will be a potential effective tool for screening the GM cry1Agene in GM crops which are widely plant in China and other developing countries.

Keywords: Insect-resistant GM crop; GM cry1A gene; Loop-mediated isothermal amplification; PCR.

 
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