CLONING AND EXPRESSION MERE GENE RELATED TO TN501 TRANSPOSON AND INVESTIGATING IT’S FUNCTION AND EFFECTS ON SWIMMING MOTILITY

	CLONING AND EXPRESSION MERE GENE RELATED TO TN501 TRANSPOSON AND INVESTIGATING IT’S FUNCTION AND EFFECTS ON SWIMMING MOTILITY
Iraqi Journal of Biotechnology
Article 1, Volume 8, Issue 2, 2009, Pages 586-594
Abstract
In this study, MerE gene related to Mer operon (Mercury resistance) from the original Tn501 transposon was amplified using PCR technique. Plasmid pUB3466 represent the cloning vector pBR322 containing the transposon Tn501 was used as a template. The PCR process was fully successful and only one band about 1.2 kbp was appeared in the gel electrophoresis. The amplified gene was cloned using pGEM-T easy vector, then transformed to a competent strain E. coli DH5α. Both of the cloning and transformation process succeeded and transformed cells were selected on selective media containing Ampicillin final concentration 100 µg/ml. Result of plasmid content analysis revealed that the transformed cell harboured one plasmid band represent the vector with MerE gene. E. coli DH5α with and without the cloned vector were used in swimming motility assay using semisolid LB media in the present and absent of HgCl2 final concentration 1µg/ml in order to study MerE function. The result showed that there was diminishment in swimming movement and the growth zone clearly decreased when HgCl2 was added. This result revealed that MerE gene may play role in transport mechanism also modified the behavior of the microorganism leading to altered motility.
Keywords
MerE gene; Mer operon; transposon; Swimming motility

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