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Anand Agricultural University, Anand

Anand Agricultural University (AAU) was established in 2004 at Anand with the support of the Government of Gujarat, Act No.(Guj 5 of 2004) dated April 29, 2004. Caved out of the erstwhile Gujarat Agricultural University (GAU), the dream institution of Sardar Vallabhbhai Patel and Dr. K. M. Munshi, the AAU was set up to provide support to the farming community in three facets namely education, research and extension activities in Agriculture, Horticulture Engineering, product Processing and Home Science. At present there seven Colleges, seventeen Research Centers and six Extension Education Institute working in nine districts of Gujarat namely Ahmedabad, Anand, Dahod, Kheda, Panchmahal, Vadodara, Mahisagar, Botad and Chhotaudepur AAU's activities have expanded to span newer commodity sectors such as soil health card, bio-diesel, medicinal plants apart from the mandatory ones like rice, maize, tobacco, vegetable crops, fruit crops, forage crops, animal breeding, nutrition and dairy products etc. the core of AAU's operating philosophy however, continues to create the partnership between the rural people and committed academic as the basic for sustainable rural development. In pursuing its various programmes AAU's overall mission is to promote sustainable growth and economic independence in rural society. AAU aims to do this through education, research and extension education. Thus, AAU works towards the empowerment of the farmers.

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Subject: Animal Genetics and Breeding × Author: Deshpande, Manisha Ramesh × End date: 2009 × Start date: 2000 × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    Study of Pituitary transcription factor, Insulin like growth factor, Leptin, Oxidized low density lipoprotein receptor 1 and Proteases inhibitor gene loci in Mehsana buffaloes
    (AAU, Anand, 2009) Deshpande, Manisha Ramesh; Rank, D. N.
    The present work was carried out to study the polymorphism in Pituitary transcription factor (PITl exon 6), Insulin like growth factor (IGFl exon 4), Leptin (exon 2), Oxidized low density lipoprotein receptor 1 (OLRl 3'UTR) and proteases inhibitor gene (PI exon 2) loci in Mehsana buffaloes through PCR-RFLP and sequencing in Mehsana buffaloes. DNA extraction was carried out by John's method from sixty blood samples of non related Mehsana buffaloes from the animals registered under progeny testing programme of Dudhsagar Research and Development Association (DURDA), Mehsana. Locus specific primers were used for PCR amplification for each of five loci. The fragment of 451bp of Pit-1 was amplified by PCR, using the primers reported by Renaville et al. (1997) and digested with restriction enzyme Hinfl. All the samples showed identical restriction pattern consisting of one site at 207 bp obtaining two fiiagments of 207bp and 244 bp. All the animals revealed monomorphic pattern with BB genotype. This result indicated no polymorphism existing in Mehsana buffalo at Pitl exon 6 locus as revealed by Hinfl RFLP. The study revealed only one allele B fixed in Mehsana buffalo with allele frequency 1.0. PCR products of representative samples were purified and cloned in pTZ57R/T vector of InsT/Aclone TM kit. Ligated recombinant vector was transformed in competent E. coli (DH5-a) cells. Recombinant plasmids were obtained and used for cycle sequencing. Sequencing revealed G—>T variation between cattle and buffalo at 283 nucleotide position. The fragment of 320 bp of IGF Iwas amplified by polymerase chain reaction, using the primers reported byDierkes e^a/.(1999) and digested with Ecol30I. On screening the IGF/Ecol30I polymorphism in Mehsana buffalo, all the animals revealed monomorphic pattern with three bands of 34, 127, and 159 bp (B allele) indicating two RE sites at 127 and 161 bp position. The investigation revealed B allele fixed in Mehsana buffaloes. As there is no polymorphism, association analysis is not warranted. PCR products of 320bp of IGF-1 (exon 4) from representative samples were purified and cloned in pTZ57R/T vector. Recombinant plasmids were obtained and used for cycle sequencing. The nucleotide sequence variation between cattle and buffalo was present at four nucleotide positions i.e. 35bp, 102bp, 132bp, 257 bp. The fragment of 331 bp of leptin gene loci was amplified by PCR, using the primers reported by Haegeman et al. (2000). Amplified products were digested with HphI and electrophoresed on 2% agarose. All the samples showed identical restriction pattern with 331 bp fiugment only. On screening the Leptia/HphI polymorphism in Mehsana buffalo one genotype AA was observed indicating allele A fixed in Mehsana buffalo. Representative sample PCR products of 331bp of leptin (exon 2) was purified and cloned in pTZ57R/T vector. Recombinant plasmids were obtained and used for cycle sequencing. The nucleotide sequence variation between cattle and buffalo was presentat six nucleotide positions i.e. 100,112,119,135,185,321. OLRl 3' UTR region was explored for Pst 1 RFLP in Mehsana buffalo. The primers reported by Khatib et al. (2006) for Bos taunts could not amplify the region in Mehsana buffaloes. Hence, new primer sets were designed using Bioinformatic tools, Primer 3.0 and Primer Express softwares (http://www.genome.wi.mit.edu/cgi bin/primer/primer3_www.cgi) on the basis of gene sequence available in the data base NW_174132.2. A 288 bp fragment of OLR gene loci was amplified by PCR, using the designed primers and digested with Pst 1. It has aPst 1 site at 215 bp and produced two fragments of 215 and 73 bp. Representative sample PCR products of 288 bp (3'UTR) was purified and cloned in pTZ57R/T vector. Recombinant plasmids were obtained and used for cycle sequencing. The nucleotide sequence variation between cattle and buffalo was present at nine nucleotide positions i.e. 85,91,116,129,151,168,171,217,240. The fragment of 448 bp of PI gene loci was amplified by PCR, using the primers reported by Khatib et al. (2005) and digested with restriction enzyme SfaNI. SfaNI digestion of PI axon 2 gene fragments revealed monomorphic pattern with appearance of three bands of 242bp, 124bp, 82 bp (B allele) in all the samples. Representative sample PCR products of 448 bp of PI (exon 2) was purified and cloned in pTZ57R/T vector. Recombinant plasmids were obtained and used for cycle sequencing. The nucleotide sequence variation between cattle and buffalo was present at ten nucleotide positions i.e. 110,141,146,170,236,257,277,306,350,396. None of the polymorphic site reported in cattle for PITl, IGFl, Leptin, OLRl and PI could be verified in buffalo. Genomic nucleotide sequences of PITl, IGFl, Leptin, OLRl and PI was submitted to Genbank of NCBI database. Bankit online sequence submission tool was used for submission of sequences to Genbank. 1. GQ385224: Bubalus bubalis POU domain class 1 transcription factor 1 (PITl) gene, exon 6 and partial cds. 2. GQ385225 : Bubalus bubalis serpin peptidase inhibitor clade A member 1 (SERPINAl) gene, partial cds. 3. GQ385226: Bubalus bubalis oxidized low density lipoprotein receptor 1 (OLRl)gene,3'UTR. 4. GQ385227: Bubalus bubalis insulin-like growth factor 1 (IGFl) gene, exon 4 and partial cds. 5. GQ385228: Bubalus bubalis leptin gene, exon 2 and partial cds.