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The BoLA class II  DQA  and  DQB  genes in crossbred cattle were studied using PCR-RFLP, cloning, and sequencing techniques. Seventy-two crossbred cattle ( Vrindavani ) were used in the current study.  Hae III and  Xba I restriction enzymes digested  DQA  exon 2-3, revealing seven ( Hae III-A-G) and three ( Xba I A-C) motifs, respectively. The BoLA- DQB  gene was analyzed using PCR-RFLP with  Pst I and  Taq I restriction enzymes, yielding five restriction motifs for each restriction enzyme ( Pst I-A-E and  Taq I-A-E). In crossbred cattle, addition, deletion, and substitutions were observed in distinct sequences, resulting in variations in overall gene length. Changes in nucleotides at positions 64–80, 110–200, and 207–264 were largely responsible for polymorphism in  DQA  exon 2. The phylogenetic analysis predicted a high degree of nucleotide and amino acid changes in DQA exon 2-3 and DQB exon 2.  DQA  genes had a nucleotide dissimilarity of 0.3–25.4 percent, while  DQB  genes had a nucleotide dissimilarity of 1.5–14.3 percent. We cloned and sequenced 20 genotypes based on PCR-RFLP of the  DQA  and  DQB  genes. The current study observed variation in the  DQA  and  DQB  genes and will serve as a foundation for future research on the BoLA  DQA  and  DQB  genes.

Characterization of BoLA class II DQA and DQB by PCR-RFLP, cloning, and sequencing reveals sequence diversity in crossbred cattle