Electrophoretic Profile Of Normal And Hydropericadium Virus-Infected Liver (Record no. 2727)

000 -LEADER
fixed length control field 03400nam a2200193Ia 4500
005 - DATE AND TIME OF LATEST TRANSACTION
control field 20150930142510.0
008 - FIXED-LENGTH DATA ELEMENTS--GENERAL INFORMATION
fixed length control field 150525s2007 xx 000 0 und d
041 ## - LANGUAGE CODE
Language code of text/sound track or separate title eng
082 ## - DEWEY DECIMAL CLASSIFICATION NUMBER
Classification number 1005,T
100 ## - MAIN ENTRY--AUTHOR NAME
Personal name Shahid Khan
110 ## - MAIN ENTRY--CORPORATE NAME
Location of meeting Prof.Dr.Irshad Hussain
245 ## - TITLE STATEMENT
Title Electrophoretic Profile Of Normal And Hydropericadium Virus-Infected Liver
260 ## - PUBLICATION, DISTRIBUTION, ETC. (IMPRINT)
Year of publication 2007
502 ## - DISSERTATION NOTE
Dissertation note Hydropericardium syndrome primarily affects the broilers between the ages of 2-7 weeks. 1he vaccine prepared from infected liver extract treated with formaldehyde is being used to protect the broilers from the disease. The current study was carried out to check the presence of immunogenic proteins in commercially available HPS vaccines Hyper immune serum was raised in 60 (3 weeks old) broiler birds using different commercially available HPS vaccine. The sera were subjected to AGPT for screening out HPS positive serum samples. The HPS infected livers were homogenized and loaded in alreadv prepared agar gel plates. The plates were incubated for 48 hours at room temperature in humid plastic box. Only 5 liver samples were found positive .The HPS autogenous vaccines,HPS positive liver samples normal liver samples were subjected to homogenization, sonication, chloroform treatment, ammonium sulphate precipitation, mixed with equal volume denaturing buffer and then boiled for 3 minutes to extract total proteins of Sample. . Large variations in the protein loading of samples in adjoining lanes lead to distortion.

Spectrophotometer determination of protein concentration assay were used to quantitate the known protein samples (bovine serum albumen is commonly used standard or this method) to standardize protein concentration. A28o were used to determine protein concentration and a calibration curve was created by plotting and performing regression analysis of A28o versus concentration of the standards and absorbance of the sample were used to determine the concentration from the calibration curve by using the formula. (concentration =Standard Factor* Dilution Factor* Optical Density.

Total protein analysis was carried out by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting technique.

A typical gel of 9% acrylamide composition was used to nicely separate polypeptides of samples to analyze the entire profile of a fraction that contains heavy and light polypeptides. Best results were obtained when 30 pi of a 20-30 mg/mi final concentrations of denatured protein sample were loaded per sample well .Each sample was repeated twice on gel. The gel was cutted at the centre vertically. 0.1% Coomassie Blue dye in 50% methanol, 10% glacial acetic acid were used to stain half of the gel for detecting protein while half of the gel was subjected to western blotting. Relative molecular weight (MW) of each protein fraction was determined by plotting a standard curve. The western blot analysis of proteins of hydropericardium syndrome virus infected liver and HPS autogenous vaccine, separated on 9% gel showed one immunogenic protein, molecular weight 15-20 kDa. However, further studies are needed to establish its immunogenic nature and feasibility for its use as vaccine.
650 ## - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical Term Department of Microbiology
700 ## - ADDED ENTRY--PERSONAL NAME
Personal name Dr. Mansur-ud-Din Ahmad
710 ## - ADDED ENTRY--CORPORATE NAME
Corporate name or jurisdiction name as entry element Faculty of Veterinary Sciences
942 ## - ADDED ENTRY ELEMENTS (KOHA)
Koha item type Thesis
Holdings
Damaged status Collection code Permanent Location Current Location Shelving location Date acquired Full call number Accession Number Koha item type
  Veterinary Science UVAS Library UVAS Library Thesis Section 2015-05-27 1005,T 1005,T Thesis


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