NDRI Quinquennium Book COVER FINAL CURVE.cdr

TheQuinquennium Golden2007-2012immunostaining positively with cytokeratin 18 and negativelywith vimentin. The BuMEC maintained the characteristicsof its functional differentiation by expression of κ-casein,κ-casein, butyrophilin and lactoferrin. BuMEC had normalgrowth properties and maintained diploid chromosomenumber (2n=50) before and after cryopreservation. Aspontaneously immortalized buffalo mammary epithelial cellline was established after 20 passages and was continuouslysubcultured for more than 60 passages without senescence.This cell line can be used as model system for studyingmammary gland functions.Differentially expressed proteins in lactatingcows having different lactation potential (Highvs low producing cows) identifiedProbable biomarkers of lactation potential in lactating cowswere identified by differential expression analysis of proteinsin mammary epithelial cells in high vs low producing Sahiwalcows, performed by differential in gel electrophoresis (DIGE)techniques within a pI range of pH 4-7 followed by MassSpectrometry. The differentially expressed proteins wereanalyzed by sophisticated decyder software in a real-timemanner. Twenty eight differentially expressed proteins havingpotential/probable role in milk yield were identified. The moststrongly regulated proteins were cytoskeletal components,calcium-binding proteins, regulators of cellular metabolismand regulators of protein stability. The identified proteins arebeing further evaluated for their biomarker potential.PCR-based method for differentiating A1 and A2beta casein containing milk developedThe DNA samples obtained from the milk were used forpresence of A1 and A2 beta-casein by using allele-specificPCR. Four different allele specific PCR based primers weredesigned and simple PCR was optimized to distinguish the A1and A2 type of beta casein. This process can be applied to milkand all products processed from that milk for identificationof A1 and A2 milk type. A mutation in the DNA sequencecoding for the beta casein protein at nucleotide position 200has resulted in the replacement of a cytidine base with adeninebase. Thus, the triplet codon affected by this change codes forhistidine (CAT) rather than for proline (CCT) at the aminoacid position 67 of the protein. Thus, the histidine at positionDIGE analysis of Sw-Hy vs Sw-Ly vs KF-Hy samples. DIGE imageswere scanned after SDS-PAGE. The gels were scanned using three laserscorresponding to Cy2, Cy3 and Cy5 wave lengths. The green colour spotsrepresesent down regulation and red colour spots represent up regulation.Sw-Hy: Sahiwal high yielding; Sw-Ly: Sahiwal low yielding; KF-Hy:Karan Fries high yielding. L: Low producer, H: High producer.The 199 bp PCR product indicates the presence of A1 beta casein allele,whereas the111bp PCR product indicates the presence of A2 beta caseinalleles.6