Sakaram, D., Niranjan, S.K., Iiumar, S., Naskar, S., Deb, S.M., Mitra, A., Sharma, A. and Sharma, D. 2010. cDNA characterization and molecular analysis of buf€alo MHC class I1 gene, DRA (Bubu-DRA). J.Appl. Anim. Res., 37: 73-76. Full cDNA of DRA gene was aniplified i n Murralz buffalo ( B u b a l u s bubalis) a.nd analysed to identify the genetic variczbtlity. Buffa,lo D R A (Bubu

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In the present study, water buffalo MHC (Bubu)-DRB cDNA was cloned and characterized. The 1022 base long-amplified cDNA product encompassed a single open reading frame of 801 bases that coded for 266 amino acids. The Bubu-DRB sequence showed maximum homology with the BoLA-DRB3*0101 allele of cattle. A total of seven amino acid residues were found to be unique for the Bubu-DRB sequence. The majority of amino acid substitutions was observed in the β1 domain. Residues associated with important functions were mostly conserved. Water buffalo DRB was phylogenetically closer to goat DRB*A.

The cDNA encoding T cell receptor-zeta (TCR-z; CD247) molecule of water buffalo (Bubalus bubalis) was isolated, cloned and sequenced in the present study. The CD247 cDNA comprised 1078 nucleotides including a 30 nucleotide 5?-untranslated region (UTR), 495 nucleotide single open reading frame (ORF) and 553 nucleotide 3?-UTR. Deduced amino acid of buffalo CD247 sequence was two residues shorter than the corresponding cattle and sheep sequences. However, ruminant-specific insertions and substitutions in intra-cytoplasmic (IC) domain were present in buffalo. Immunoreceptor tyrosine-based activation motifs (ITAMs), the important motifs for TCR signalling, were totally conserved among ruminants including buffalo. The 3?-UTR region of the buffalo CD247 was highly homologous to the corresponding region in the cattle sequence and showed lack of polymorphism after polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis using HaeIII and MseI restriction enzymes in buffalo population. Phylogenetically, buffalo sequence was closer to cattle sequence under the ruminant's lineage. The conserved nature of this gene ensures TCR integrity which is vital for induction of optimal and efficient immune response.

Sexed semen can be used to produce offspring of the desired sex, and to produce replacement heifers from genetically superior cows. In the Indian dairy industry, this revolutionary technique is in its budding stage. The objective of this study was to assess quantitative and qualitative parameters of a) bovine cryopreserved sexed (for X sperm cells) (XS) spermatozoa produced through two different major commercial semen sorting techniques, and b) unsexed (US) spermatozoa of Bos indicus and Bos taurus breeds. By real-time PCR, it was found that the percentage of X chromosome bearing spermatozoa ranged from 62.88%±0.56 to 71.21±0.93 in US semen samples of four different cattle breeds: Holstein Friesian (HF), Jersey, Gir and Sahiwal. The corresponding values in semen sorted through flow cytometry technique ranged from 99.02% ±3.36 to 99.99%±4.56 in the above four cattle breeds; while the percentage of XS semen obtained through the decapitation of Y chromosome following sorting by flow cytometry ranged from 95.92% ±1.83 to 97.99%±2.74. Qualitatively, US sperm samples were found significantly (P<0.01) superior with high semen motility (65.98% ± 0.23), acrosomal integrity (75.28% ± 0.05), live spermatozoa (67.83% ± 0.42), and less sperm abnormalities (13.82% ± 0.02) than sexed semen samples. This is the first study to compare the quantitative and qualitative parameters of XS and US semen samples of major cattle breeds in India and adds significantly to the knowledge on semen sexing in Indian dairy industry.