Study of differential gene expression in response to exogenous administration of bovine somatotropin hormone in Indian buffalo (Bubalus bubalis)
Ramani, Umed (2007) Study of differential gene expression in response to exogenous administration of bovine somatotropin hormone in Indian buffalo (Bubalus bubalis). Masters thesis, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand, Gujarat, India.
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Technological developments in a variety of scientific and engineering disciplines will be needed to support the growing world population, which is expected to double in the next 40 years. It has been estimated that the supply of food required to adequately meet human nutritional needs over the next 40 years is quantitatively equal to the amount of food previously produced throughout the history of humankind. To meet this need, it will be essential that scientists continue to develop new technologies that increase efficiency of food production. With respect to livestock industry, administration of exogenous Somatotropin (ST) is one that increases the food output (meat or milk) per unit of feed resource input. Bovine somatotropin is a growth hormone found in cattle. The word bovine refers to cattle, and the word somatotropin refers to the name of the hormone. They are natural substances that affect the way the body operates. Bovine somatotropin, abbreviated as bST, is a protein hormone produced in cattle by the pituitary gland located at the base of the animal’s brain. Now, the new science of systems biology makes it possible to work with DNA, the part of a cell that contains the genetic information for an animal or a plant. Scientists have determined which gene in cattle controls or codes for the production of bST. They have removed this gene from cattle and inserted it into a bacterium called Escherichia coli. This bacterium, which is found in the intestinal tract of humans and animals, acts like a tiny factory and produces large amounts of bST in controlled laboratory conditions. The bST produced by the bacteria is purified and then injected into cattle to increase milk synthesis and meat production. The present study was undertaken with the objectives to identify the up/down regulation of m-RNA and candidate genes involved in the process of increase in milk synthesis by sequence characterizing the differentially expressed m-RNA in udder tissue collected at 0, 48 and 96 hours of rbST treatment from Indian buffalo through DDRT-PCR. Total mRNA was extracted using TRIZOL based methods. First strand cDNA synthesis was prepared using Omniscript reverse transcriptase kit than go for PCR of cDNA with three anchored primers (HT11m) and four arbitrary (Osteo-poietin, HAP01, HAP03 and HAP25). Differentially displayed (DD) band was eluted from PAGE. DD bands were sequenced and analysis part of differentially expressed ESTs was carried out using different bioinformatics tools like blastn, blastX, available at NCBI. Fifty DD bands were sequenced, 22 EST found differentially displayed and 28 were up regulated as intensity of band in gel was increase from 0 to 96 hrs of rbST treatment and appear only in treated animal tissues, respectively. Out of 50 ESTs, 18 ESTs shown location on chromosome no. 24 where 1, 6, 3, 1, 1, 2, 1, 2, 1, 2 and one ESTs were shown location on 6, 7, 13, 14, 16, 17, 18, 19, 21, 22 and 27, respectively. Five ESTs had partial match but 5 transcripts were not showing any significant sequence similarity with any chromosome. Out of 50 sequences, 9 EST sequences were showing similarity with specific protein gene, 41 sequences were not matching with any protein, meaning, it may not be reported in database of bovine tissues. While out of 9, 7 were showing similarity with specific protein raboconnectin and 2 with TSC1 protein. Out of 50 EST sequences, 9 were showing match with katanin protein which is located on intronic region of chromosome no. 24. Out of 50 ESTs, 12 ESTs and 19 ESTs were showing match with different tissues while 38 and 31 ESTs were not showing any significant sequence similarity in blastn and Bos taurus ESTs division that means still there is no report of these ESTs in Bos taurus or any other database. In the transcriptome world, 5 ESTs out of 50 ESTs were not specified any match in any database so it is first time reported in the buffalo mammary tissues which are novel transcript found to be responsible for increasing milk synthesis. From this study it can be concluded that large number of novel transcripts as well as ESTs with known functions are involved in increased milk production. In systems biology, further investigation is required for sequencing of full length cDNA and expression of cDNA sequence in appropriate host, interaction among these differentially expressed transcripts.
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