Cloning and sequencing of partial regions in chromosome II of B. abortus and chromosome I of B. melitensis

Abstract

In the present study, PCR amplified products of partial alk B gene (498 bp) and that of 731 bp products of B. abortus and B. melitensis strains respectively were analysed. Further, the 498 bp product of B. abortus was ligated into pGEM-T Easy vector and the recombinant plasmid after RE digestion with EcoRI released the insert and sequenced. Upon comparison of the nucleotide sequence of B. abortus S 99 strain with those of reference sequences from USA, it was found that, there was 100% homology with all the three sequences. Similarly, the PCR amplified product of B. melitensis 16M strain (731 bp) was similarly cloned and the nucleotide sequences were aligned with two reference sequences from USA and one from Belgium. B. abortus and B. melitensis strains showed 100 and 98.8% similarity respectively with the reference sequences available, thus confirming the purity of the strains being used in the diagnostic kits.