ISOLATION OF ROTAVIRUS FROM DIARRHEA IN CHILDREN AND DEVELOPMENT OF DNA VACCINE
Usha Tiwari*, Nitin Sharma, Pragya Pandey and Manoj Kalakoti
ABSTRACT
Rotavirus present in faecal samples was extracted. Rotavirus genome consists of 11 double stranded RNA (ds RNA) segments enclosed in a double-shelled capsid. The cDNA of VP7 gene was synthesized and amplified by RT-PCR, thus PCRproduct had a total length of 1039 nucleotides. It was ligated in pVAX1 vector. Prepared the competent cell to transformation of ligated product in E. coli DH5?. LB agar plates were inoculated and large numbers of colonies were picked from the overnight grown transformants. The individual colonies were inoculated in fresh kanamycin(50 ?g/ml) containing LB broth and allowed to grow for large scale production for further experiments, i.e. for expression and immunological studies.
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