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Exploration of Genetic Polymorpphisms and Differential Expression Analysis of Bovine Alpha-Lactalbumin and Osteopontin Genes Involved in Milk Composition

Thesis Info

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External Link

Author

Manzoor, Dr. Sidra

Program

PhD

Institute

University of Veterinary and Animal Sciences

City

Lahore

Province

Punjab

Country

Pakistan

Thesis Completing Year

2017

Thesis Completion Status

Completed

Subject

Molecular Biology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/13163/1/Thesis%20Sidra%2c%2020%20sep%2c%202017.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726160493

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Economically important traits of dairy animals are usually controlled by a large number of genes. The identification of the single nucleotide polymorphisms in potential genes has been associated with economically important traits. During lactation, mammary epithelial cells produced large amounts of specific milk proteins. Due to the expression sites, physiological properties and chromosomal localization, LALBA and SPP1 genes might be considered as candidate genes for milk composition in buffalo. Alpha-lactalbumin (LALBA) gene has been reported to be highly transcribed in transition and peak phase while late lactation exhibited its decline with progressive rise in SPP1 expression. This project was designed to investigate the effect of single nucleotide polymorphism that influencing the gene expression thus modulates the milk protein content in Nili Ravi. Samples of unrelated Nili-Ravi buffalo were collected from two Government, Buffalo Research Institute, Pattoki, and Livestock Production and Research Institute (LPRI) Bahadarnagar Okara, livestock farms. Milk samples were collected at 15, 90 and 250 days lactation for expression analysis. The genomic DNA was extracted by using the standard Phenol Chloroform Isoamyl alcohol (PCI) protocol. Specific set of primers was designed for the amplification of the LALBA and SPP1 genes. The amplified PCR products were sequenced for the identification of SNPs. To determine the differential expression of bovine LALBA and SPP1 genes, RNA was isolated from milk samples using the TRIzol reagent and converted it into cDNA. Taqman probes were used that are specifically designed to detect and target the DNA sequence. Five intronic polym orphic sites were identified in LALBA while exonic regions exhibited a complete homology with reference sequence. Additionally, eleven polymorphisms were identified in bovine SPP1 gene, six were in coding region and five were found in intronic portion of the gene. The analysis and correlation of all identified polymorphism was done by using SNPs data analysis software “SNPator”. Results obtained from expression study was stored in in-build software of Real Time PCR and Cycle threshold (Ct) values of LALBA and SPP1 mRNA were compared in individuals of Nili-Ravi buffalo to determine the variation in expression levels. The LALBA gene expression was observed highest in transition phase with a gradual decrease of expression in mid and late lactation. The sample, NR-5, was observed highly expressed (79.30) while NR-2 with low expression (19.28) for alpha lactalbumin in early lactation. The change in LALBA regulation at same stage was considered due to genetic variation of the respective animal. While the SPP1 gene expression was observed with the highest values in peak lactation and remains elevated in late lactation. NR-4 has the highest (72.27) expression among all mastitis free healthy animals while NR-2 was observed with low expression. Thus, the identified SNPs might be used as genetic marker for milk production traits. Gene expression patterns may also help us to understand the molecular mechanisms of bovine LALBA and SPP1 genes influencing milk composition. However, the expression of both genes was considered in a correlation with other genes involved in milk production pathway. Also, the mutational effects of other milk proteins might be involved in determining the expression pattern of both genes in selected animals. Therefore, further studies are likely to explore the regulation of milk protein genes and their translational efficiency during the course of lactation in dairy animals.
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حاجی اقبال احمد

حاجی اقبال احمد
افسوس ہے ۱۶،۱۷/اپریل کی درمیانی شب میں ہمارے علاقے اوردہلی کے مشہور ومعروف صاحب خیر حاجی اقبال احمد صاحب ۷۲ سال کی عمر میں رحلت فرما گئے۔ مرحوم۱۹۴۷ء کے انقلاب سے قبل پھاٹک حبش خاں میں انڈے،مرغی کی تجارت کرتے تھے۔انقلاب کے بعد جامع مسجد کے علاقے میں آگئے اوریہی کاروبار اورمچھلی کاکاروبار وسیع پیمانے پر کرنے لگے۔ ہرشہر میں بڑے بڑے تاجر ہوتے ہیں، حاجی صاحب بھی ایک بڑے کاروباری تھے لیکن ان کی غیر معمولی خصوصیت یہ تھی کہ اوّل درجے کے صاحب خیر تھے، امورخیر کی جستجو اور تلاش میں رہتے تھے اوراجتماعی اور ملی کاموں میں ذوق وشوق سے حصہ لیتے تھے، یہی وجہ ہے کہ دوردور تک ان کی شہرت تھی۔رمضان المبارک میں مدارس عربیہ کے سینکڑوں سفیر ان کے یہاں آتے تھے اورمرحوم بڑے حوصلے سے ان مدرسوں اور دینی درسگاہوں کی خدمت کرتے تھے۔ سفیروں کے ہجوم اورکثرت کی وجہ سے گزشتہ کئی سال سے یہ معمول بنالیا تھا کہ رمضان المبارک کی۲۱تاریخ سے زکاۃ کی تقسیم شروع کرتے تھے اورپھر آخر تک یہ سلسلہ قائم رہتا تھا۔مسجد مچھلی دالان میں چندہ لینے والوں کی لائنیں لگ جاتی تھیں اورحاجی صاحب صبر و برداشت بلکہ خندہ پیشانی سے ان سب کی مدد کرتے تھے، پتے لکھے ہوئے سینکڑوں منی آڈر فارم علیٰحدہ آتے تھے جو عید کے بعد روانہ کیے جاتے تھے۔ بیواؤں اورنادار شریف گھرانوں میں بیٹھی ہوئی نوجوان لڑکیوں کی شادیوں میں امداد کرناان کا محبوب مشغلہ تھا۔ ویران اوراُجڑی ہوئی مسجدوں کی تعمیر اورآبادی میں والہانہ انداز سے حصہ لیتے تھے، رنگ روڈپر شاہ بڑے کی حسین وجمیل اور وسیع مسجد ان کی حرارت ایمانی کی زبردست یادگار ہے۔ وزیراعظم کی سمادھیوں کے درمیان اس خوبصورت مسجد کے سبک میناروں کی عجیب شان نظرآتی ہے، انقلاب سے پہلے یہ مسجدزیادہ آباد نہیں تھی اب...

M.H. Panhwar as a Historian

The scope of this article is to find out the strengths and weaknesses of M.H. Panhwar’s historical work and to compare it with other scholars of Sindh. He is the initiator of scientific history writing of Sindh and the only historian who tried to uncover the face behind the social organisation of Sindh. According to him, ‘History is the history of production, control over the means of production and production to its final distribution’. The history is more about people than that of the ruling class as people are makers of history. He had vision to analyze things in their correct historical perspective and, in this context, he took support of archeology, as it is the source of scientific truth. Writing history with data acquired through carbon testing is a new phenomenon in Sindh, which was locally introduced by M. H Panhwar.1 M. H. Panhwar, therefore, was one of the pioneers in giving a new meaning to history by including both ruling elites and the common folk, progressive science, crop technology, forest, wildlife, etc.2 He was the first to throw light on areas which were ignored by other historians. According to him, history written before him is full of the accounts of the rulers’ role in relation to their nobles and foes, battles they fought, attacks, palace plotting, mausoleums, folklore, fiction and so on. Accounts regarding contemporary climate, environments, diversion of rivers, courses and the resultant ruination of irrigation systems, migrations, shifting of people, occupations, social life of the populace etc. Found little room in written histories. For instance changes in production and its means, administration and justice, agriculture, land, animal husbandry, irrigation sources and methods of mining, metallurgy, industry and industrial goods, matrimonial institutions, family and children, houses and interiors, art, architecture, archaeology, personal appearance, dresses, ornaments, foods and drinks, taxes, coins and currency, science and technology, foreign contacts, international trades and traders, routes of trades, religious beliefs, philosophy, hygiene, medicine and doctors, superstitions, common citizens, economic conditions, historical geography of bygone ages, classes of work and their relation with one another, household life, customs, entertainments, pastimes, leisure, attitude of man to nature, languages, literature, literary contacts with outside word, learning and thoughts, status of women and children in affluence and extreme poverty.3 Hence, in real sense M.H Panhwar was scholar of Haig’s Raverty’s and Lambrick’s calibre.

Improving the Performance of Maize Zea Mays L. by Silicon Application under Drought Condition

Water shortage on our globe has very deleterious effects on food security. Judicious use of water for crops seems very wise in the current scenario of water scarcity. Huge research work is required to investigate how can we grow crops with less amount of water? To determine the solution for such kinds of problems this research project was designed. The research project entitled, “Improving the performance of maize (Zea mays L.) by silicon application under drought condition” was carried out at Postgraduate Agricultural Research Station (PARS), University of Agriculture, Faisalabad, Pakistan. In the first pot experiment screening of maize hybrids (P-33H25, S-8441, FH-985, ICI-8288, YH-1921, R-3333, FH-963, FH-810) against drought was carried out. Moisture levels of 100%, 80% and 60% field capacity were maintained. From this experiment the most tolerant (P-33H25) and most sensitive (FH-810) maize hybrids against drought were screened out. In the second pot experiment four silicon levels (0, 50, 100 and 150 mg/kg) were optimized under moisture levels of 100% and 60% field capacity maintained. The optimized silicon level (100 mg/kg) was selected for further study. The third pot experiment was conducted to investigate the improvement in drought stressed maize hybrids by exogenous application of silicon. The two selected hybrids from the first experiment were sown under two moisture regimes (100% and 60% field capacity). In this experiment silicon level i.e. 100 mg/kg (optimized in experiment II) was used. Drought was imposed two weeks after emergence and maintained. The fourth experiment was conducted under field condition with two selected maize hybrids (P-33H25 and FH- 810) and two irrigation regimes (100% and 60% FC) from the 1 st experiment and two silicon levels (0 mg/kg and 100 mg/kg) from 2 nd experiment. In this experiment moisture application was maintained at 100 or 60% field capacity. In this experiment the maize crop was sown for two years 2010 and 2011 in both seasons i.e. spring and autumn. The data regarding morphological, physiological and biochemical parameters were recorded by following standard procedures. The analysis of data from the first experiment showed that mild and severe drought had deleterious effects on various morphological and physiological parameters. However, root fresh and dry weight enhanced under mild drought while decreased under severe water limited condition. On the basis of dry matter production the most drought tolerant (P-33H25) and drought sensitive (FH-810) maize hybrids were screened out. In the second experiment best suited silicon level (100 mg/kg) was optimized on the basis of dry matter production. In the third experiment drought severely affected the morphological, physiological and biochemical attributes of maize hybrids. However, silicon application improved the various agronomic attributes, yield and yield components. The same study was conducted in field as that of third experiment to investigate the effect of silicon on maize hybrids grown under water limited condition. Significant effects of silicon application were observed on various agronomic, physiological, yield and yield related parameters. However, further research is required to investigate the silicon mediated drought tolerance in maize hybrids.