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Authors: Khirbat, Richa
Advisor: Sangwan, M.L.
Title: Polymorphism in lactoferrin gene and its association with mastitis in Murrah buffaloes
Publisher: LUVAS
Language: en
Type: Thesis
Keywords: Lactoferrin, Murrah, PCR-RFLP, PCR-SSCP, Sequencing
Abstract: The present investigation was undertaken to study the genetic polymorphism in lactoferrin gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and single strand conformational polymorphism (PCR-SSCP) technique along with nucleotide sequencing and its association with mastitis in 40 Murrah buffaloes (including 20 mastitic animals). Genomic DNA was isolated from randomly selected lactating buffalo maintained at Govt. livestock farm, Hisar, Animal farm and Veterinary clinics, LLRUVAS, Hisar, by phenol chloform isoamyl alcohol method. DNA quality and quantity was checked by agarose gel electrophoresis. PCR was standardized to amplify partial promoter region (370bp) and exon 15 (261bp) of lactoferrin gene using published and specific self designed (NCBI pick primer) primers. When 261 bp PCR products of exon 15 of lactoferrin gene were digested with Taqα I and Nru I restriction endonucleases, it detected single genotype viz. xx. Only monomorphism was observed with all restriction enzymes under study. When SSCP analysis of 370 bp PCR products of promoter region of lactoferrin gene was done it revealed 5 SNPs between healthy and mastitis animals. In comparison of the sequence with reference sequence Bubalus bubalis (EF650854.1) and Seyfert sequence,some important SNPs at positions +35, -19 and -156 in accordance with literature were observed. The T–C transition (+35) was observed in the 5′ UTR of exon1, 4 bp upstream of the start codon which could play a crucial role in the regulation of gene expression. No significant association of lactoferrin gene with mastitis was found. However there was significant (P<0.05) difference in somatic cell count of healthy (0.41x105 ± 0.6x104) and mastitis animals (26.35x105 ± 40.9x104). In phylogenetic tree, the lactoferrin gene (exon 15 and promoter region) of Bubalus bubalis are found more closely related to Bos taurus than to Capra hircus
Subject: Animal Biotechnology
Theme: Polymorphism in lactoferrin gene and its association with mastitis in Murrah buffaloes
These Type: M.V.Sc.
Issue Date: 2012
Appears in Collections:Thesis

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