Preparation method of envelope protein A gene vaccine for fowl cholera
A technology of outer membrane protein and fowl cholera, applied in gene therapy, pharmaceutical formula, genetic material components, etc., can solve problems such as easy recurrence, side effects, and unsatisfactory immune effect
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[0052] Step one, primer design
[0053] Two primers P1 and P2 were designed to amplify the outer membrane protein A gene of Fowl Cholera according to the genome sequence of Fowl Cholera:
[0054] P1: 5’-GCGGTACCatgaaaaaaacagcaattgc-3’;
[0055] P2: 5’-CGCGCGAATTCTTATTTGTTACCTTTAACAGCG-3’;
[0056] Step 2: PCR amplification and purification of the outer membrane protein A gene of Fowl Cholera
[0057] 1) Add 1μl each of primer P1 and primer P2 to a 0.2ml PCR reaction tube, then add 0.5μl of DNA polymerase, 2.5μl of polymerase buffer, 3μl of four dideoxyribonucleotide mixtures, Avian Pasteurella 1μl of the genome and 16μl of water, and then put the PCR reaction tube containing the above reagents into the PCR amplification machine, and carry out the PCR reaction. The PCR reaction procedure: first carry out the pre-denaturation at 94oC for 5 minutes, and enter the circulation process after the pre-denaturation is over , The procedure of each cycle is: denaturation at 94oC, time is 1 minut...
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