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In the present study, antisense primers were designed from 7 differentially displayed expressed sequence tags (ESTs) previously identified at our laboratory. Total RNA was isolated from the mammary gland tissue of lactating buffalo to carry out 5' rapid amplification cDNA ends (RACE) PCR. 5' RACE PCR amplicons, ranging from 161 bp to 336 bp, were cloned and sequenced. These 5' RACE sequences were analyzed using the blastn (Megablast) bioinformatics tool available from NCBI. Of these 7, only one 5' mRNA sequence did not show a match in the available database and another 1 failed to show a match with any specific gene due to the unavailability of buffalo genome information. Two other 5' mRNA sequences showed 82% to 100% homology with the 18S rRNA gene and the remaining 3 showed homology to the Talin-1, β casein, and 28S rRNA genes. Full length cDNA were synthesized using primers designed from 5' mRNA sequences and ranged from 345 bp to 1030 bp. Finally, we conclude that the ESTs identified may act as markers for lactogenesis in other animals. [ABSTRACT FROM AUTHOR]

Background: Human leukocyte antigen (HLA) is comprised of a highly polymorphic set of genes which determines the histocompatibility of organ transplantation. The present study was undertaken to identify HLA class I and class II allele, genotype and haplotype frequencies in renal transplant recipients and donors from West Central India., Materials and Methods: HLA typing was carried out using Polymerase Chain Reaction-Sequence Specific Primer in 552 live related and unrelated renal transplant recipients and donors., Results: The most frequent HLA class I and class II alleles and their frequencies in recipients were HLA-AFNx0101 (0.1685) and AFNx0102 (0.1649), HLA-BFNx0135 (0.1322), and HLA-DR beta 1 (DRB 1)FNx0115 (0.2192), whereas in donors, these were HLA-AFNx0102 (0.1848) and AFNx0101 (0.1667), HLA-BFNx0135 (0.1359), and HLA-DRB1FNx0115 (0.2409). The two-locus haplotype statistical analysis revealed HLA-AFNx0102-B61 as the most common haplotype with the frequency of 0.0487 and 0.0510 in recipients and donors, respectively. Further, among the three locus haplotypes HLA-AFNx0133-BFNx0144-DRB1FNx0107 and HLA-AFNx0102-BFNx0161-DRB1FNx0115 were the most common haplotypes with frequencies 0.0362 and 0.0326, respectively in recipients and 0.0236 and 0.0323, respectively in donors. Genotype frequency revealed a high prevalence of genotype HLA-AFNx0102/AFNx0124 in recipients (0.058) compared to donors (0.0109) whereas low prevalence of HLA-AFNx0101/AFNx0102 in recipients (0.0435) than in donors (0.0797). The phylogenetic and principal component analysis of HLA allele and haplotype frequency distribution revealed genetic similarities of various ethnic groups. Further, case control analysis provides preliminary evidence of association of HLA-A genotype (P < 0.05) with renal failure., Conclusion: This study will be helpful in suitable donor search besides providing valuable information for population genetics and HLA disease association analysis.