Author(s):
1. Areeba Yousaf:
Department of Biochemistry, Institute of Biochemistry, Biotechnology and Bioinformatics, The Islamia University of Bahawalpur, Bahawalpur, Pakistan
2. Mirza Imran Shahzad:
Department of Biochemistry, Institute of Biochemistry, Biotechnology and Bioinformatics, The Islamia University of Bahawalpur, Bahawalpur, Pakistan
Abstract:
Tuberculosis (TB) is a contagious and infectious disease which is caused by Mycobacterium tuberculosis (M.tb). TB remains a lethal pathogenic disease in history. It is still an uncontrolled disease because of the endless drug resistance, unavailability of an effective vaccine, coinfection with HIV/AIDS as well as the absence of timely diagnostic techniques. TB is also considered a disease in underdeveloped countries where sufficient health facilities are deficient. Novel vaccination regimens against M.tb are urgently needed. The aim of the present study is to design an effective DNA vaccine construct against TB and to study its immunization effects on mice models. For this purpose Mycobacterium gene, InhA/Rv1484 was selected. The gene was amplified by using gene specific primers. Mammalian expression vector pVAX1 was transformed into competent cells DH5a i.e. strain of E.coli. Both the insert and vector were ligated after their double restriction digestion using endonuclease enzymes XbaI and HindIII. Additionally, vector pVAX1 was dephosphorylated before ligation to avoid its self-ligation. The ligated product was transformed into competent cells. The colonies were picked and grown for plasmid extraction. The successful cloning was confirmed through PCR of extracted plasmid with gene specific and pVAX1 specific primers. Restriction digestion of the construct was also performed to confirm the cloning. Sequence analysis of the construct was also done to verify the desired DNA vaccine construct. Later, this construct was extracted following an endotoxin-free plasmid extraction procedure. The DNA vaccine was inoculated in the mice model. Other groups of mice were also vaccinated with BCG and pVAX1.The cocktail of DNA vaccines was prepared with otherM.tb DNA vaccines and inoculated in a mice group. The DNA vaccine was also used as the prime boost dose in BCG vaccinated mice model. The tests on the blood/serum of mice model were performed for immunization studies. The antibody titer will be confirmed through multiplex microbead assay. The efficacy of the BCG vaccine is also enhanced with DNA vaccine construct in the form of prime-boost. The glycerol stock of the clone was prepared and saved at -80? to be used in challenge studies.
Page(s):
51-51
DOI:
DOI not available
Published:
Journal: Abstract Book on Global Science Technology and Management Conference, Volume: 0, Issue: 0, Year: 2023
Keywords:
DNA vaccine
,
Mycobacterium Gene InhaRv1484
,
Mice Model
References:
References are not available for this document.
Citations
Citations are not available for this document.