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Researcher Buragohain, Lukumoni
Advisor Nanda, Trilok
Source KrishiKosh-Indian National Agricultural Research System
Content type Text
Educational Degree Master of Science (M.Sc.)
Publisher LUVAS
File Format PDF
Language English
Subject Domain (in DDC) Technology ♦ Agriculture & related technologies ♦ Animal husbandry
Subject Keyword Proteomics ♦ 2-d Gel Electrophoresis ♦ Densitomeric ♦ Mass Spectrometry Expasy ♦ Mx2 ♦ Animal Biotechnology ♦ Serum Proteomics and Mx2 Gene Expression Analysis In The Peripheral Blood of Early Pregnant Buffalo
Abstract Modern proteomics technique offers an exciting opportunity in solving some of the most challenging problems of livestock sector, including development of diagnostics for early pregnancy. In the present study, blood samples obtained at weekly intervals from early pregnant (Day 0 to Day 35 post-AI) buffalo heifers, and on days 0, 7 and 14 from non-pregnant cyclic buffalo heifers. The sera samples were subjected to depletion of high abundant proteins followed by 2-dimensional gel electrophoresis and densitometric analysis, which revealed the presence of at least 10 2-D gel spots exhibiting upregulation or specific appearance at a specific stage during early buffalo pregnancy, except for the spots correlating with the high abundant proteins’ location. Mass Spectrometry analysis and comparison with ExPASy and NCBI databases matched 10 of these spots with known proteins, but with varying degrees of confidence in terms of Mascot score and the species. Although high abundant proteins were depleted before 2-D electrophoresis, yet in some of the picked spots isoforms of common abundant proteins viz. Serum Albumin, IgG, Serrotransferrin, Complement molecules, were found. Some of these identified spots appear promising pregnancy bio-markers, especially synaptojanin-1, apolipoprotein A-II, Keratin 10 and Serum Amyloid A, which are documented to have role in embryogenesis and early pregnancy. The same blood samples were also used for RNA extraction to synthesised cDNA which was later used as template in Real Time PCR for profiling of the relative gene expression of MX2 transcript in pregnant and non-pregnant animals with respect to endogenous control which in present study was GAPDH. In case of pregnant buffaloes expression of MX2 increases significantly on day 14, 21 and 28 which ranges from 16 to 30 fold change as compared to other days of pregnancy and non pregnancy. For the first time the MX2 protein was estimated in serum of pregnant and non pregnant buffalo hefiers at the said days. The MX2 protein concentration was also significantly higher on 14, 21 and 28 days of pregnancy with respect to others of pregnancy and non pregnancy. Keeping the current study in view, it seems that Proteomics and expression study of MX2 gene would open a new window towards the development of rapid early diagnostic tool for buffalo which might have field application in near future.
Educational Use Research
Education Level UG and PG
Learning Resource Type Thesis
Size (in Bytes) 5.89 MB