Journal of Pure and Applied MicrobiologyVol. 4 No. 2

Screening and Cloning of Partial cry1Ac Gene Fragment from an Indigenous Isolate of Bacillus thuringiensis

R. Manikandan*, V. Kannan, C. Muthukumar and P. Logeshwaran

Centre for Plant Molecular Biology, Tamil Nadu Agricultural University, Coimbatore - 641 003, India.

Received on 02 April 2010 and accepted on 14 May 2010

 

ABSTRACT

PCR is a useful technique for quick and simultaneous screening of Bacillus thuringiensis strains for classification and prediction of insecticidal activities. This experiment has been contacted by screening of cry1Ac gene from new isolates Bt. Two indigenous isolates of Bt were used as a template for screening of cry1Ac genes by mixtures of degenerate primer and specific primer of cry1Ac genes. Two new isolates of Bt, T17, 14r1 showed amplification with its cry1Ac gene. But among these two strains, 14r1 strain was showed variation during their amplification (~925bp). The partial cry1Ac gene fragment amplified from Bt isolates, 14r1 and cloned in E. coli vector. The recombinants clones of cry1Ac gene (~925bp) were also showed variation in their amplification.

Keywords : Bacillus thuringiensis, cry1Ac gene, degenerate primers, cloning.