Identification of differentially expressed genes during different physiological stages of mammary gland in Indian buffalo (Bubalus bubalis)
Nandasana, Kamlesh (2007) Identification of differentially expressed genes during different physiological stages of mammary gland in Indian buffalo (Bubalus bubalis). Masters thesis, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand.
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A key challenge to be met by biotechnologist, genetician and dairy scientists is to understand which genes control the composition of milk, how these genes are regulated and how they might be manipulated to enhance both, the manufacturing and health properties of dairy products. The mammary gland is subject to dramatic adaptations throughout the lifespan of a female. Structural changes of the mammary gland are most evident during the dry period and they prepare the udder for the onset of lactation. After calving, the mammary gland begins to produce many components (fat, protein, lactose) and secretes them in milk. All physical adaptations during the dry period and throughout lactation must occur through coordinated increases or decreases in the expression of many genes. The present study was undertaken with the objective to identify the differentially expressed candidate gene for milk synthesis through DDRT-PCR in udder tissues of dry, heifer, pregnant and lactating buffaloes. Out of four, one sample each from udder tissue of dry, heifer, pregnant and lactating buffaloes were taken for the purpose of the study. The analysis part of differential ESTs was carried out using different bioinformatics tools like blastn and blastX, available at NCBI. Using three anchored as well as five arbitrary primer a total of 33 differential bands were eluted and analysed. Out of 33 EST sequences, 20 ESTs were differentially expressed and 13 ESTs were upregulated. Nine ESTs were located on chromosome no. 5, 7 ESTs on chromosome no. 24, 2 ESTs on chromosome no. 13, 2 ESTs on chromosome no. 9 and one EST on each of chromosome no. 1, 2, 4, 7, 11, 14, 17, 28, and 29, while location of 4 ESTs on chromosome was not specified. Nineteen sequences were showing similarity with ESTs of specific protein gene, 9 sequences were showing similarity with ESTs with no specific protein gene, 4 sequences were showing similarity only with chromosome, while one sequence was showing no similarity with any database. Out of 19 ESTs of specific protein, 8 sequences were showing similarity with triose phosphate isomerase mRNA, 4 sequences were showing similarity with rabconnectin-3 beta mRNA, 2 sequences were showing similarity with α- lactalbumin mRNA, 2 sequences were showing similarity with katanin mRNA, one sequence was showing similarity with KIAA0776 mRNA, one sequence was showing similarity with mannosyl α-1,3-glycoprotein β-1,4-Nacetylglucosaminyltransferase, isozyme B (MGAT4B), mRNA and one sequence was showing similarity with butyrate response factor 2, mRNA. Rabconnectin-3 beta, triose phosphate isomerase and KIAA0776 was differentially expressed in mammary tissue of heifer, pregnant and lactating buffalo respectively. Protien katanin was also differentially expressed in lactating buffalo mammary tissue. However, α-lactalbumin and butyrate response factor 2 was highly upregulated in mammary tissue of lactating buffalo while mannosyl α-1,3-glycoprotein – β-1,4- N-acetylglucosaminyltransferase, isozyme B (MGAT4B) was highly upregulated in mammary tissue of pregnant buffalo. Out of 14 ESTs, which were not specifying any protein, 3, 1 and 1 ESTs were differentially expressed in mammary tissue of pregnant, lactating and dry buffalo, respectively, while 4, 4 and 1 ESTs were highly upregulated in mammary tissue of pregnant, lactating and heifer buffalo, respectively. Four ESTs were showing similarity with Bos taurus ESTs while 9 ESTs were novel. EST A3GL- 1.25 was showing no similarity with any database, also considered as novel.
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