Cloning of amylase gene in E.coli-Bacteroides pGFK114.1 shuttle vector and conjugation between Ecoli strains for possible application in rumen digestive system.

To increasing the efficiency of production in livestock industry could use recombinant DNA technology. Recently progress in gene transfer system، genetic modifying of ruminal bacteria with ability of hydrolytic enzyme production has been possible. Resulting this technology reduce of disadvantages of enzyme supplements as well as increase of nutrient flow in rumen. Since transformation in rumen bacteria has low efficiency therefore conjugation could be effective for moving genetic elements. In more previous studies E. coli as a facultative anaerobe bacteria has used because could grow in rumen. The present study، an amylase gene previously clone in pET28a and amplified by polymerase chain reaction then subcloned in shuttle vector pGFK114. 1 following transformed in E. coli DH5α. Conjugation was performed between two E. coli DH5α and TG1 donors containing pGFK114. 1 and pRK231 helper plasmid respectively and one recipient E. coliBL21 DE3 using pattern of digestive system. We have successfully transformed cloned amylase gene to recipient bacteria such rumen environment as reported before.