Isolation, PCR Amplification and Cloning of Heat Shock Protein Gene from Salivary Glands of Hyalomma dromedarii Ticks from Camelus dromedarius

Abstract

A molecular study was carried out to isolate the heat shock protein gene of Hyalomma dromedarii ticks of camels, which play important role in their survival under harsh environmental conditions. Engorged adult H. dromedarii ticks were collected from camel herds in the Bikaner district of Rajasthan. Total genomic DNA and RNA was isolated from the salivary glands of ticks. Primers were designed for amplification of heat shock protein gene from H. dromedarii by using base sequence of Haemaphysalis longicomis. The heat shock protein gene of H. dromedarii was successfully amplified from genomic DNA and cDNA and was identified on the basis of its size in agarose gel electrophoresis as 560 bp. The amplicon of expected size was purified from the 1% low melting agarose gel. DNA fragment of interest was then ligated to the pGEM-T Easy vector and ligated mixture was transformed into Escherichia coli JM109 strains for cloning. Screening of recombinants was done by restriction enzyme digestion of plasmid DNA and by colony PCR for quick screening of plasmid insert directly from E. coli colonies in the presence of insert specific primers.