Publications (27)24.4 Total impact
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Article: Microsatellite (GT)(n) polymorphism at 3'UTR of SLC11A1 influences the expression of brucella LPS induced MCP1 mRNA in buffalo peripheral blood mononuclear cells.
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ABSTRACT: A (GT)(n) microsatellite polymorphism at 3'UTR of SLC11A1(solute carrier family 11A1) is associated with the natural resistance to bovine brucellosis. A pleiotropic effect of SLC11A1 on other candidate genes influencing the host resistance including monocyte chemotactic/chemoattractant protein 1 (MCP1) is also hypothesized. In the present study, we report the cloning and characterization of the complete coding sequence of bubaline (bu) MCP1 and its tissue distribution at the transcript level. The buMCP1 exhibited as high as 99% and >80% of sequence identities with the bovine and other domestic animal species homologues. The buMCP1 mRNA was abundant across the different tissues: most abundant in liver and mammary gland, moderate in ovary, skeletal muscle and testis, and least in uterus. Further, quantitative real-time PCR (RTqPCR) analysis revealed that PBMCs carrying so called resistant GT(13) allele produced more MCP1 mRNA endogenously as well as when induced with brucella LPS suggesting the pleiotropic roles of SLC11A1 in conferring resistance against the intracellular pathogens particularly against brucellosis. However, the underlying molecular mechanisms by which 3'UTR SLC11A1 concomitantly increases the production of chemokines like MCP1 are yet to be investigated.Veterinary Immunology and Immunopathology 01/2013; · 2.08 Impact Factor -
Article: Characterization of insulin like growth factor-1 (IGF-1) partial gene in mithun
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ABSTRACT: Insulin like growth factor -1 (IGF-1), a protein hormone similar to insulin in molecular structure (Blundell et al.1978), is produced by liver and target tissues and the hepatocytes are main site for its synthesis (Schwander et al.1983). IGF-1 is considered as a major hormone, which is involved intricately in the growth of animals. The gene for IGF-1 extends over > 45 kilo base pairs, and contains 5 exons which are separated by 4 large non-coding introns ranging in size from 1.9–40 kb (Rotwein et al.1986). Exon 1 and 2 encode portions of the signal peptide whereas exon 3, 4 and 5 code for mature IGF1 molecule. Although there are several reports regarding the polymorphism in IGF-1 gene in different species, viz. cattle (Ge et al 1997), swamp buffaloes (Dierkes et al.1999), goats and buffaloes (Mishra 2005) and dogs (Sutter et al. 2007) are available but no reports are there on mithun -an important beef animal of NE India, which plays a significant role in the economic, social and cultural life of tribal people. Due to high meat value of this animal, the growth is the most desirable parameter for improvement. The present study is an attempt to characterize and ascertain polymorphism in the exon 5 of IGF1 gene of mithun. Unrelated mithun (90) maintained at National Research Centre on Mithun, Medziphema and Porba farm, Nagaland from all 4 types, viz. Arunachalee, Nagamee, Manipuri, Mizoram were selected for this study. 15 ml of intravenous blood was collected from each animal in sterile 15 ml polypropylene centrifuge tube containing 0.5 ml of 2.7% EDTA as anticoagulant. DNA isolation was done by phenol-chloroform extraction method (Sambrook et al. 2001) and the precipitated DNA was dissolved in 200 µl of TE buffer. The quality of DNA was checked by performing agarose gel electrophoresis. The purity and concentration were evaluated by spectrophotometry. The sample that showed an OD ratio (OD 260 /OD 280) in the range of 1.7–1.9 were assessed to be of good quality. Concentrations of DNA samples were in the range of 16–20 µg/ml of blood. A set of primers (5' ATG TCA CTT TTT CTC GCT TAT T3' as forward primer and 5' CAT GCA TTT GTG GCT CTT G 3' as reverse primer) was designed on the basis of cattle genomic sequences (AF210383S5), to amplify a fragment (396 bp) corresponding to part of intron 4 (27 bp), exon 5 (60 bp) and part of 3′ flank (309 bp) of IGF1 gene. The stock was reconstituted by DNAse free milipore water to the tune of 300 pmoles//µl. The working primer solution was then prepared by further 10 fold dilution of stock solution to have a final concentration of 30 pmole/µl. The reaction mixture and PCR programme were optimized to achieve the satisfactory level of amplification in a final volume of 25 µl containing 1µl of genomic DNA (80–100 ng), 2.5 µl of 10 X PCR buffer (1.5 mM), 2.5 µl of dNTP mix (0.2 mM), 1.5 µl of MgCl 2 (1.5 mM), 1 µl each of forward primer and reverse (30 pmol/µl), 0.2 µl of Taq DNA polymerase (5U/µl). Samples were amplified for 35 cycles with initial denaturation at 94°C for 3 min, cyclic denaturation 94°C for 45 sec, cyclic annealing at 60°C for 45 sec, cyclic extension at 72°C for 45 sec followed by final extension at 72°C for 10 min. It was digested with MspI (HpaII) restriction enzyme, which cuts at the recognition site of 5′…C↓CGG…3′. The RE digestion was carried out in 15 µl reaction mixture carrying 7 µl of autoclaved triple distilled water, 2 µl of 10X assay buffer for RE, 1 µl of RE enzyme (10U/µl) and 15 µl of PCR product. The incubation was carried out at 37°C for overnight for complete digestion. Restriction fragments were resolved in 2.5% agarose gel stained with ethedium bromide in 1X TBE buffer and visualized under UV light. The amplicon of IGF-1 gene was eluted and sequenced. The obtained sequence was then aligned with other available sequences of different species from GenBank (www.ncbi.nlm.nih.gov). A portion of IGF-1 gene corresponding to 27 bp of intron 4, complete exon 5 having 60 bp length and 309 bp of 3lanking region constituted the amplicon of 396 bp length and its digestion with MspI (C↓CGG) produced 2 bands i.e.The Indian journal of animal sciences 02/2012; 82:89-91. · 0.12 Impact Factor -
Article: Molecular characterization of MHC-DRB cDNA in water buffalo (Bubalus bubalis).
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ABSTRACT: 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 β(1) domain. Residues associated with important functions were mostly conserved. Water buffalo DRB was phylogenetically closer to goat DRB*A.Genetics and Molecular Biology 01/2012; 35(1):95-8. · 0.63 Impact Factor -
Article: Analysis of genetic variations of complete TM4 of buffalo (Bubalus bubalis) Slc11A1 gene
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ABSTRACT: To explore polymorphisms at exon V–VII, encompassing complete TM4 and part of TM3 and TM5 of buffalo SLC11A1 gene, 150 animals belonging to three breeds of buffalo (Bubalus bubalis) viz., Murrah, Surti and Mehsana were genotyped by PCR-RFLP using five restriction enzymes (REs) (Alu I, Taq I, Rsa I, Sac I and Bse GI). PCR–RFLP revealed monomorphic restriction pattern among the three buffalo breeds. Although, sequence comparison revealed no variations at nucleotide level among the buffalo breeds under study, however, two synonymous substitutions at amino acid position 145 (A→C transversion) and 151 (T→C transition) of TM3 were observed compared to cattle sequence (NM-174652). No changes in TM4 and synonymous substitutions at TM3 reflect the conserve nature of this region specially TM4 in buffalo. Future studies may be directed to explore polymorphisms throughout the entire buffalo SLC11A1 gene and to ascertain their suitability as potential genetic marker for resistance against various diseases.Journal of Applied Animal Research 12/2011; 39(4):324-327. · 0.40 Impact Factor -
Article: Genetic characterisation of buffalo MHC (Bubu)-DQB cDNA molecule
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ABSTRACT: A 771 nucleotide long cDNA molecule corresponding to major histocompatibility complex (MHC)-DQB gene was amplified, cloned and sequenced from water buffalo (Bubalus bubalis). Open reading frame of the Bubu-DQB sequence was short by three nucleotides from the DQB genes of the other ruminants. Nucleotide similarity of Bubu-DQB was highest with cattle DQB alleles (84–97%), followed by goat (88.8%) and sheep (87.5%). The sequence comparison revealed that the DQB gene was highly variable in buffalo particularly, in exon 2 (β1 domain). A total of 45 amino acid substitutions were identified in Bubu-DQB compared to cattle DQB*0101 sequence, with a maximum (27) in β1 domain. The residues involved in antigen binding and heterodimer formation were found to be different in buffalo DQB sequence compared to other species. Phylogenetic study showed that the Bubu-DQB has evolved prior to the diversification of common DQB alleles in ruminants. Our study revealed the high genetic variation in the DQB gene in buffalo, which might have generated to recognise species specific antigens.Journal of Applied Animal Research 06/2011; 39(2):136-138. · 0.40 Impact Factor -
Article: Molecular Characterization and SNP Detection of CD14 Gene of Crossbred Cattle.
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ABSTRACT: CD14 is an important molecule for innate immunity that can act against a wide range of pathogens. The present paper has characterized CD14 gene of crossbred (CB) cattle (Bos indicus×Bos taurus). Cloning and sequence analysis of CD14 cDNA revealed 1119 nucleotide long open reading frame encoding 373 amino acids protein and 20 amino acids signal peptide. CB cattle CD14 gene exhibited a high percentage of nucleotide identity (59.3-98.1%) with the corresponding mammalian homologs. Cattle and buffalo appear to have diverged from a common ancestor in phylogenetic analysis. 25 SNPs with 17 amino acid changes were newly reported and the site for mutational hot-spot was detected in CB cattle CD14 gene. Non-synonymous substitutions exceeding synonymous substitutions indicate the evolution of this protein through positive selection among domestic animals. Predicted protein structures obtained from deduced amino acid sequence indicated CB cattle CD14 molecule to be a receptor with horse shoe-shaped structure. The sites for LPS binding, LPS signalling, leucine-rich repeats, putative N-linked glycosylation, O-linked glycosylation, glycosyl phosphatidyl inositol anchor, disulphide bridges, alpha helix, beta strand, leucine rich nuclear export signal, leucine zipper and domain linker were predicted. Most of leucine and cysteine residues remain conserved across the species.Molecular biology international. 01/2011; 2011:507346. -
Article: Allelic diversity at MHC class II DQ loci in buffalo (Bubalus bubalis): evidence for duplication.
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ABSTRACT: The genetic diversity of MHC class II DQ genes was investigated in riverine buffalo (Bubalus bubalis) by PCR-RFLP and sequencing. Highly variable regions (exons 2-3) of DQ genes were amplified from 152 buffaloes and genotyped by PCR-RFLP. Alleles identified by differential restriction patterns were sequenced for the characterization. PCR-RFLP was a rapid method to discriminate between DQA1 and duplicated DQA2 genes in buffalo, however, the method appeared to be inadequate for determining the more complicated DQB genotypes. A total of 7 and 10 alleles were identified for DQA and DQB loci, respectively. Nucleotide as well as amino acid variations among DQ alleles particularly at peptide binding regions were high. Such variations were as expected higher in DQB than DQA alleles. The phylogenetic analysis for both genes revealed the grouping of alleles into two major sub-groups with higher genetic divergence. High divergence among DQ allelic families and the isolation of two diverse DQA and DQB sequences from individual samples indicated duplication of DQ loci was similar in buffalo to other ruminants.Veterinary Immunology and Immunopathology 12/2010; 138(3):206-12. · 2.08 Impact Factor -
Article: PCR-SSCP of Serum Lysozyme Gene (Exon-III) in Riverine Buffalo and Its Association with Lysozyme Activity and Somatic Cell Count
Asian Australasian Journal of Animal Sciences 07/2010; 23(8-23(8)):993. · 0.58 Impact Factor -
Article: Nucleotide variability in partial insulin like growth factor 1 (IGF-1) gene of mithun
The Indian journal of animal sciences 09/2009; 79(9):939-941. · 0.12 Impact Factor -
Article: Cloning and characterization of DGAT1 gene of Riverine buffaloFull Length Research Paper
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ABSTRACT: The present study was carried out to characterize the DGAT1 gene of Riverine buffalo. Total RNA was extracted from the mammary tissue of buffalo and DGAT1cDNA were synthesized by RT-PCR, then cloned using pDRIVE cloning vector and sequenced. The sequencing revealed that the size of DGAT1 gene was 1470 bp with GC content of 62.30%. The gene encoded for 489 amino acid precursors and that it possessed 32 amino acids signal peptide. The similarity of buffalo DGAT1 mRNA sequence with that of cattle, pig, monkey, human, mice and rat were determined as 98.4, 90.7, 85.4, 85.0, 77.4 and 77.1%, respectively. Phylogenetic tree constructed from the derived DGAT1 protein sequences of 15 different species illustrated a unique branches for mammals, fly, nematode and plants. Among mammals, cattle and buffalo grouped together, whereas swine formed another group in the same branch. Four motifs were predicted in buffalo DGAT1 peptide sequence, one N-linked glycosylation site (246th position), two putative tyrosine phosphorylation site (316 and 261), one putative diacylglycerol binding site (382–392 amino acid position) and a conserved domain MBOAT (membrane bound acyl transferase from 150 to 474 amino acids) with a histidine as an active residue.07/2009; 19(3):177-184. -
Article: Cloning and characterization of αs2-casein gene of Riverine buffaloFull Length Research Paper
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ABSTRACT: The present study was carried out to characterize the αs2-casein gene in Riverine buffalo. Total RNA was extracted from the mammary tissue of buffalo and αs2-casein cDNA were synthesized by RT-PCR, then cloned using pDRIVE-cloning vector and sequenced. The sequencing revealed that the size of αs2-casein was 669 bp with GC content of 41.11%. The gene encoded for 222 amino acid precursors and that it possessed 15 amino acids signal peptide. The similarity of buffalo αs2-casein mRNA sequence with that of cattle, sheep, goat, pig and camel were estimated as 97.9, 93.6, 93.4, 73.5 and 73.0%, respectively. In the phylogenetic trees, constructed from the data of the αs2-casein mRNA sequences as well as protein sequences, it has been observed that the cattle and buffalo were in the same group whereas sheep and goat formed another group. The camel and swine were placed in two separate groups.07/2009; 17(6):458-464. -
Article: Isolation of two cDNAs encoding MHC-DQA1 and -DQA2 from the water buffalo, Bubalus bubalis.
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ABSTRACT: In the present study, we explored structural and functional variations and possible duplication of the major histocompatibility complex (MHC)-DQA gene in water buffalo (Bubalus bubalis). Two cDNA sequences, amplified from one individual water buffalo, were designated as Bubu-DQA1 (DQA*0101) and -DQA2 (DQA*2001). The percentage of nucleotide and amino acid similarity between Bubu-DQA1 and -DQA2 revealed that these sequences display more similarity to alleles of respective DQA1 and DQA2 genes from other ruminant species than to each other. The phylogenetic analysis also revealed a considerably larger genetic distance between these two genes than between homologous genes from other species. The larger genetic distance between DQA*0101 and DQA*2001, and the presence of different bovine DQA putative locus specific amino acid motifs, suggests these sequences are non-allelic. This finding is consistent with DQA gene duplication in other ruminants.Veterinary Immunology and Immunopathology 03/2009; 130(3-4):268-71. · 2.08 Impact Factor -
Article: Identification of novel allelic variants of integrin beta 2 (ITGB2) gene and screening for Bubaline leukocyte adhesion deficiency syndrome in Indian water buffaloes (Bubalus bubalis).
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ABSTRACT: A fragment of 570 bp corresponding to exon 5 and 6 of integrin beta 2 (ITGB2) gene was amplified for screening D128G mutation in one hundred and fifty two buffaloes (Bubalus bubalis) which causes bovine leukocyte adhesion deficiency syndrome (BLAD) in cattle, as well as to ascertain polymorphism. TaqI PCR-RFLP revealed no such mutation thus indicating the absence of bubaline leukocyte adhesion deficiency (BuLAD) allele in animals under study. However, the polymorphism studies using MspI restriction enzyme revealed two genotypic patterns viz. AA pattern (bands of 293, 141, 105, and 31 bp) and BB pattern (bands of 293, 105, 77, 64, and 31 bp). The sequences of A and B alleles were submitted to the GenBank (EU853307 and AY821799).Animal Biotechnology 02/2009; 20(3):156-60. · 0.93 Impact Factor -
Article: Lack of Polymorphism in Partial Insulin Like Growth Factor 1 (IGF1) and Insulin Like Growth Factor Binding Protein 3 (IGFBP3) Genes of Mithun
Journal of Applied Animal Research 01/2009; 36(1):41-44. · 0.40 Impact Factor -
Article: Cloning and characterization of DGAT1 gene of Riverine buffalo.
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ABSTRACT: The present study was carried out to characterize the DGAT1 gene of Riverine buffalo. Total RNA was extracted from the mammary tissue of buffalo and DGAT1cDNA were synthesized by RT-PCR, then cloned using pDRIVE cloning vector and sequenced. The sequencing revealed that the size of DGAT1 gene was 1470 bp with GC content of 62.30%. The gene encoded for 489 amino acid precursors and that it possessed 32 amino acids signal peptide. The similarity of buffalo DGAT1 mRNA sequence with that of cattle, pig, monkey, human, mice and rat were determined as 98.4, 90.7, 85.4, 85.0, 77.4 and 77.1%, respectively. Phylogenetic tree constructed from the derived DGAT1 protein sequences of 15 different species illustrated a unique branches for mammals, fly, nematode and plants. Among mammals, cattle and buffalo grouped together, whereas swine formed another group in the same branch. Four motifs were predicted in buffalo DGAT1 peptide sequence, one N-linked glycosylation site (246th position), two putative tyrosine phosphorylation site (316 and 261), one putative diacylglycerol binding site (382-392 amino acid position) and a conserved domain MBOAT (membrane bound acyl transferase from 150 to 474 amino acids) with a histidine as an active residue.DNA Sequence 07/2008; 19(3):177-84. · 0.75 Impact Factor -
Article: Association of microsatellite (GT)n polymorphism at 3'UTR of NRAMP1 with the macrophage function following challenge with Brucella LPS in buffalo (Bubalus bubalis).
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ABSTRACT: Brucella abortus is a facultative intracellular pathogen that survives and replicates in host macrophages. Hence, macrophage function plays an important role in influencing natural resistance/susceptibility to intracellular pathogen. The natural resistance associated macrophage protein 1 (NRAMP1; erstwhile referred as Ity/Lsh/Bcg), a transmembrane protein, regulates activity of macrophages against intracellular pathogens. In bovine, natural resistance to brucellosis is significantly associated with (GT)13 allelic variant of microsatellite locus at 3' untranslated region (3'UTR) of the NRAMP1 gene. In the present study we screened 65 Murrah breed of buffalo (Bubalus bubalis) to identify polymorphism at 3'UTR of NRAMP1 gene and evaluate the association of these polymorphisms with the macrophage function. Four allelic variants (viz., GT13, GT14, GT15 and GT16) were identified. Majority of the buffaloes were of either homozygous (GT)14/(GT)14 or heterozygous (GT)14/(GT)15 with (GT)14 allele occurring most frequently (62%). For association study, non-vaccinated and serologically negative animals were divided into three genotypic groups: group 1 (n=2) comprising animals of homozygous (GT)13 genotype, whereas, group 2 (n=4) and group 3 (n=6) consisted animals of heterozygous [(GT)13/(GT)n, where n not equal 13] and non-(GT)13 [(GT)n/(GT)n, where n not equal 13] genotype, respectively. Macrophages, after maturation, were challenged with Brucella LPS to assay the macrophage function in terms of H2O2 and NO production. The (GT)13 allele, either in homozygoous {(GT)13/(GT)13} or heterozygous {(GT)13/(GT)n, where n=14, 15 or 16}, was significantly (p<0.01) associated with increased production of H2O2 and NO. In this manuscript, for the first time, we have identified (GT)13 allelic variant and demonstrated its significant association with the improved macrophage function in buffalo.Veterinary Microbiology 05/2008; 129(1-2):188-96. · 3.33 Impact Factor -
Article: Cloning and characterization of alpha(s2)-casein gene of Riverine buffalo.
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ABSTRACT: The present study was carried out to characterize the alpha(s2)-casein gene in Riverine buffalo. Total RNA was extracted from the mammary tissue of buffalo and alpha(s2)-casein cDNA were synthesized by RT-PCR, then cloned using pDRIVE-cloning vector and sequenced. The sequencing revealed that the size of alpha(s2)-casein was 669 bp with GC content of 41.11%. The gene encoded for 222 amino acid precursors and that it possessed 15 amino acids signal peptide. The similarity of buffalo alpha(s2)-casein mRNA sequence with that of cattle, sheep, goat, pig and camel were estimated as 97.9, 93.6, 93.4, 73.5 and 73.0%, respectively. In the phylogenetic trees, constructed from the data of the alpha(s2)-casein mRNA sequences as well as protein sequences, it has been observed that the cattle and buffalo were in the same group whereas sheep and goat formed another group. The camel and swine were placed in two separate groups.DNA Sequence 01/2007; 17(6):458-64. · 0.75 Impact Factor -
Article: Genetic variants of beta-lactoglobulin gene and its association with milk composition traits in riverine buffalo.
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ABSTRACT: A study was carried out to determine genetic variants of beta-lactoglobulin gene and to explore associations between these and milk composition traits in riverine buffalo. Single strand conformation polymorphism was employed to detect the genetic variants of the gene. Two fragments of this gene i.e. 119 bp of exon I and 400 bp spanning exon IV and intron IV were included in the study. For 119 bp fragment, three alleles namely, A, B and C were observed in all the buffalo breeds whereas four alleles (A, B, C and D) were detected for 400 bp fragment. The frequency distribution of alleles was different in different breeds of buffaloes for both the fragments. For exon I fragment, the milk composition traits such as total SNF, protein, solid, fat and whey protein yield were found to be significantly (P<0.05) associated with genotypes in Murrah and Bhadawari buffalo whereas in Mehsana breed genotypes were significantly (P<0.05) co-related with total SNF, solid and fat yield. Genotypes of 400 bp fragment, only total fat yield in Mehsana buffalo was found to be significantly (P<0.05) associated with genotypes.J Dairy Res 12/2006; 73(4):499-503. · 1.57 Impact Factor -
Article: Molecular characterization of the interferon-tau gene of the mithun (Bos frontalis).
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ABSTRACT: The mithun (Bos frontalis) not only remains one of the most neglected ungulate species due to its remote range, but also has been identified as a vulnerable species due to its declining population. Augmenting its reproductive efficiency could be a strategy for reversing its population decline. Considering the importance of interferon-tau (IFNT) as a primary signal in establishing maternal recognition of pregnancy (MRP), the present study was undertaken to characterize the IFNT gene of the mithun. A 588 bp mithun IFNT (mitIFNT) gene was PCR amplified using genomic DNA as the template. Its nucleotide sequence comprised an entire open reading frame of 585 bp encoding a 195 amino acid pre-protein. In nucleotide sequence, the mitIFNT gene was more than 85% similar to the homologous genes of domestic and wild ruminant species characterized to date. However, phylogenetic analysis placed mitIFNT into a clade containing IFNT of the red deer, but not IFNTs of cow, sheep, or goats, or other wild ruminant species. Our characterization of mitIFNT represents the first complete sequence of any gene from the mithun.ZOOLOGICAL SCIENCE 08/2006; 23(7):607-11. · 0.95 Impact Factor -
Article: DRB3.2 gene polymorphism and its association with pashmina production in Changthangi goat. :
International Journal of Immunogenetics. 01/2006; 33:271-276..
Top co-authors
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Institutions
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2003–2012
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Indian Veterinary Research Institute
- Division of Animal Genetics
Bareilly, Uttar Pradesh, India
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2005
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National Dairy Research Institute
Karnāl, Haryana, India
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