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Study On Polymorphism Of Promoter Region Of Bovine Lactoferrin Gene And Its Relation With Mastitis In Nili Ravi Buffalo

By: Muhammad Asim (2012-VA-636) | Dr. Sehrish Firyal.
Contributor(s): Dr. Muhammad Yasir Zahoor | Dr. Muhammad Tayyab.
Material type: materialTypeLabelBookPublisher: 2017Description: 79p.Subject(s): Molecular Biology and BiotechnologyDDC classification: 2923-T Dissertation note: Dairy animals in Pakistan and worldwide are facing most persistent and economically affecting disease mastitis. Only a healthy buffalo can produce good quality milk of physiologically normal composition. Mammary gland inflammation due to mastitis badly affects the quantity and quality of milk and this causes big loss to dairy industry. Even province Punjab bears economic losses of 240 million per annum due to mastitis. Susceptibility and resistance to mastitis is affected by the variation in immunity genes. Among immunity genes Lactoferrin (LF) have important role in immune defense system and perform antibacterial, antiviral and anti-inflammatory function. LF is found in most of body fluids like, milk, blood, tear, saliva, bile and mucous. Polymorphism in promoter region of Lactoferrin gene is associated with mastitis susceptibility and resistance. For screening of the mastitis susceptibility and resistance of dairy buffaloes, LF is a potential candidate gene. The present study was designed for the identification of polymorphism in LF gene associated with mastitis. Blood samples from 20 Nili Ravi buffalos having clinical and subclinical mastitis were selected. Blood samples of normal Nili Ravi buffalos were also collected. DNA was extracted; specific primers for amplification of LF gene were designed with Primer-3 software by using already reported sequence on NCBI. Amplification of LF gene performed by PCR and sequenced the amplicon. A comparative analysis of sequence result was performed by using NCBI BLAST. BioEdit software was used to perform multiple sequence alignment. Comparison analysis of LF gene promoter region shows multiple mutations in in clinical and subclinical as compare to reported sequence (accession no. EF650854) at NCBI. While normal samples sequence results are similar to reference data. These results show LF is a candidate gene for mastitis resistance.
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Dairy animals in Pakistan and worldwide are facing most persistent and economically affecting
disease mastitis. Only a healthy buffalo can produce good quality milk of physiologically normal
composition. Mammary gland inflammation due to mastitis badly affects the quantity and quality
of milk and this causes big loss to dairy industry. Even province Punjab bears economic losses of
240 million per annum due to mastitis.
Susceptibility and resistance to mastitis is affected by the variation in immunity genes. Among
immunity genes Lactoferrin (LF) have important role in immune defense system and perform
antibacterial, antiviral and anti-inflammatory function. LF is found in most of body fluids like,
milk, blood, tear, saliva, bile and mucous. Polymorphism in promoter region of Lactoferrin gene
is associated with mastitis susceptibility and resistance. For screening of the mastitis
susceptibility and resistance of dairy buffaloes, LF is a potential candidate gene.
The present study was designed for the identification of polymorphism in LF gene associated
with mastitis. Blood samples from 20 Nili Ravi buffalos having clinical and subclinical mastitis
were selected. Blood samples of normal Nili Ravi buffalos were also collected. DNA was
extracted; specific primers for amplification of LF gene were designed with Primer-3 software
by using already reported sequence on NCBI.
Amplification of LF gene performed by PCR and sequenced the amplicon. A comparative
analysis of sequence result was performed by using NCBI BLAST. BioEdit software was used to
perform multiple sequence alignment. Comparison analysis of LF gene promoter region shows
multiple mutations in in clinical and subclinical as compare to reported sequence (accession no.
EF650854) at NCBI. While normal samples sequence results are similar to reference data. These
results show LF is a candidate gene for mastitis resistance.

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