Molecular Charaterization Of Ampk Gene Of Pakistan Buffalo
Material type: Book ; Format:
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Publisher: 2011 Dissertation note: Pakistan is an agriculture country and its economy is mainly dependent on agriculture, agriculture products. Livestock has been playing an important role in the economy of the country. Livestock sector contributed approximately 51.8 percent of the agriculture value added and 11.3 percent to national GDP. Buffalo which is known as black gold of Pakistan is famous for its largest milk production in the world. A better understanding of the genetic control of energy metabolism in farm animals can have far-reaching implications for molecular breeding programs. It can allow the implementation of knowledge-based breeding to increase feed efficiency and to improve meat quality. In addition, because of the high degree of evolutionary conservation of these genes, the information gained about the genetic control of animal nutrition can be extrapolated back to questions about human nutritional genomics and disease. This study was performed to discover the single nucleotide polymorphism at AMP-activated protein kinase (AMPK) gene in Nilli Ravi and Kundi Buffalo and their possible association with milk production. As AMPK is a sensor of energy metabolism so genetic variations in AMPK gene may also have effect the feed utilizing efficiency of animals. Buffalo is popular for utilizing low quality roughages in a better way. Buffaloes are popular in the world for high fat content and low cholesterol content as compare to cattle. A total of 128 single nucleotide polymorphisms were discovered at AMPK gene in Nilli-Ravi and Kundi Buffalo. Out of which 10 are in exonic region and 118 are in Intronic region. Most of the SNPs are Intronic it also shows that AMPK is highly conserved as it has been shown by many studies. The Intronic SNPs may have role in regulation of AMPK gene. Forty-six SNPs were discovered in Intronic region of A1 subunit of AMPK gene. Out of these 46 SNPs. Forty-four SNPs are same in both Nilli-Ravi&Kundi buffalo.
Two SNPs found at position 11908 and 12217 was present only in Kundi buffalo. These two SNPs can be used for breed characterization of Nilli-Ravi&Kundi buffalo. The numbers of SNPs discovered in exonic region are 6. These all SNPs are non-synonymous mutations and changes amino acids at position 23333 from Histidine>Tyrosine, at 23387 from Glutamic acid>Lysine, at 23402 from Valine>Isoleucine, at 23426 from Ser>Pro, at 23489 from Stop codon>Arg and at 23612 from Ala>Thr. Forty SNPs were discovered in Intronic region of A2 subunit of AMPK gene. Out of these 43 SNPs 28 are same in both Nilli-Ravi & Kundi buffalo. SNPs at positions 71371, 71382, 71383, 71396, 71558, 42736, 42766, 42881, 41661, 41900 and 42021 are only present in Kundi buffalo while SNPs at position 70900, 71613, 42935 and 42944 are present only in Nilli-Ravi buffalo. These SNPs can also be used for breed characterization of Nilli-Ravi and Kundi buffalo. The B1 subunit of AMPK gene has 21 SNPs in Intronic region, which is common, both in Nilli-Ravi and Kundi buffalo. These polymorphisms may have role in regulation of AMPK gene. The SNPs found in exonic region are 3 which are all non-synonymous mutations and changes amino acids at position 4362 from Histidine>Tyrosine and at positions 8193, 8195 from Glycine>Serine. All exonic SNPs are non-synonymous mutations, which show that it will change the function of protein and might be associated with milk production and feeding efficiency in Nilli-Ravi & Kundi buffalo. This study is an example of candidate gene approach to find some novel variations at population level. It is the first study conducted for Molecular Characterization of AMPK gene in Buffalo. The only way to associate these polymorphisms to the trait under consideration (energy metabolism) by back tracing the sampling groups. This study is first in finding some molecular markers for energy metabolism in Nilli-Ravi and Kundi buffalo that can be used for future selection and breeding programs. More the population will be diversified for the trait and showing trends of heterozygosity, better will be the chances of selection of animals with suitable genetic makeup.
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Genetic And Evolutionary Characterization Of Pakistani Pigeons And Parrots Through Mitochondrial D-
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Publisher: 2013 Dissertation note: Abstract
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Differential Expression And Mutation Analysis Of Heat Shock Proteins (Hsps) And Tumor Suppressor Gene (P53) In Differemt Cancer Types of Pakistani Dogs and Cats
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Publisher: 2014 Dissertation note: Abstract
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Bioconversion Of Agricultural Waste To Alginate By Azotobacter Vinelandii Using Fermentation
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Publisher: 2015 Dissertation note: Alginate is an exopolysaccharide composed of varying ratios of β-D mannuronic acid and its C5 epimer α-L-guluronic acid linked together by β-1,4 - glycosidic bond. It has wide range of industrial applications particularly in food sector as a viscosifier, stabilizer, thickener, emulsifier, gelling and water binding agent. Commercial alginate is extracted from brown algae but due to variation in composition of biopolymer isolated from species of different locations, there is growing interest in bacterial alginate.
At present two strains of bacteria are reported to produce alginate, Pseudomonas and Azotobacter. Hence present study was designed to produce alginate by Azotobacter vinelandii utilizing cheap substrates to save the foreign exchange. To achieve the goal, different physio-chemical parameters were optimized to have hyper-production of alginate through submerged fermentation. Different agricultural wastes like wheat bran, rice polishing and molasses were utilized as substrates through fermentation with Azotobacter vinelandii.On fermentation of 7.5% (w/v) wheat bran by A.vinelandii, maximum alginate production (5.21 g/L) was observed at 48 hours of incubation time with 6% (v/v) inoculum size, pH 7.0, 300C and agitation speed of 200 rpm. Addition of different optimum levels of ionic salts i.e. 1.5% CaCl2 and 2% MgSO4. 7H2O in the growth medium gave significantly (P< 0.05) higher quantity of alginate (6.08 g/L) where as addition of KH2PO4 and NaCl reduced the yield of alginate. Among different nitrogen sources tested, 2% corn steep liquor resulted significantly (P<0.05) higher yield of alginate (7.46 g/L).
The bacterial strain was improved by exposure to physical (UV irradiation) and chemical mutagens (Nitrous acid and ethidium bromide) to obtain more than 90% killing. The survivors were screened for hyper-production of alginate against the wild strain of A.vinelandii using pre-optimized conditions. The highest alginate production (13.8 g/L) was obtained by the ethidium bromide treated strain (EtBr-02). The mutant strain was used for optimization of fermentation parameters. The maximum concentration of alginate (15.61 g/L) was obtained by utilizing 10% (w/v) wheat bran, 8% (v/v) inoculum at 48 hours of incubation, pH 7.0, 300C and an agitation speed of 200 rpm. Inclusion of 2.5% cornsteep liquor raised the alginate concentration to 15.8 g/L.
Batch fermenter studies were carried out in 2 L fermenter with working volume of 1.5 L using the mutant strain A.vinelandii, EtBr-02. Optimization of process parameters like agitation, aeration and pH in the fermenter showed that maximum alginate (16.8 g/L) was achieved at 300 rpm, 2.5 vvm aeration and controlled pH condition at 32 hours of incubation time.
The alginate produced was identified by FTIR spectrum after precipitation. The purity of alginate was estimated by HPLC against the standard alginic acid from Sigma-Aldrich and was found to be 98% pure. The alginate produced was used at 3% concentration for immobilization of yeast cells. Immobilized and free cells were compared for ethanol production using 10% sucrose as the carbon source in fermentation medium. The maximum amount of ethanol obtained was from free cells i.e. 38 g/L whereas immobilized cells produced 32.5 g/L ethanol. The advantage of immobilization is that beads can be reused in eight sequential fermentation cycles of 10 h each. Thus a cheap and practical bioprocess of alginate production was developed, that can be exploited commercially to save foreign exchange.
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