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Tissue Specific Esterase Expression in Indigenous Chicken of Bangladesh

R. M. Shahjahan
Laboratory of Animal Genetics and Molecular Biology, Department of Zoology, University of Dhaka, Dhaka-1000, Bangladesh

K. A. Muid
Laboratory of Animal Genetics and Molecular Biology, Department of Zoology, University of Dhaka, Dhaka-1000, Bangladesh

Q. S. Akter
Reproductive Biotechnology Laboratory, Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

K. M. A. Tareq
Reproductive Biotechnology Laboratory, Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

R. A. Begum
Laboratory of Animal Genetics and Molecular Biology, Department of Zoology, University of Dhaka, Dhaka-1000, Bangladesh

R. Begum
Laboratory of Animal Genetics and Molecular Biology, Department of Zoology, University of Dhaka, Dhaka-1000, Bangladesh

M. A. M. Y. Khandoker
Reproductive Biotechnology Laboratory, Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

The present investigation was done in the esterase gene expression analysis of the Red Jungle fowl Gallus gallus of Bangladesh utilizing the polyacrylamide gel electrophoresis (PAGE) technique. Each allele produces a biochemical message that helps to produce various protein phenotypes those were expressed in the form of bands on the polyacrylamide gel bed. Altogether four different esterase bands were recorded in both Red Jungle (RJ) fowl and Naked Neck (NN) fowl of Bangladesh. These were denoted as Est-1 (2), Est-2(1), Est-3 (21) and Est-4 (13) based on the relative mobility on the gel. Among them Est-2 is very common in almost all tissues. Comparing the esterase in both genetic groups, their expression intensity even the relative mobility provided a unique feature that they were similar. The investigation indicates that esterase gene between the Red Jungle fowl and Naked Neck fowl of Bangladesh has a close relationship.

Keywords: Esterase, Gel electrophoresis, Naked Neck fowl, Red Jungle fowl

Location: Bandarban District of Bangladesh

Start Date: 00-00-2008

End Date: 00-00-2008

Program Area: Animal Health and Management

Commodity: Chicken

Objectives:

1. To understand the genetic status of Red Jungle (RJ) fowl of Bangladesh in terms of esterase variability.

Methodology:

The whole carcass of the RJ fowl was collected from Bandarban District of Bangladesh. According to the requirement the tissue of forebrain, midbrain, hindbrain, heart, liver, testis, oesophagus, gizzard, crop, proventiculous, small intestine, large intestine, pectoral muscle, pelvic muscle, lung, eye, kidney, pancreas, body muscle and blood were collected. Separate special sterile bags were used for carrying the samples to avoid the contamination. The samples were then transported to the laboratory in an insulated box with ice to maintain the temperature around 4-6°C. In the laboratory, each and every sample was taken separately and kept in deep freezer at -80°c until further processing. Different tissues of the RJ fowl were taken in a measure nearly about 0.0159 gm. Each sample was taken to an eppendrof tube and squashed in 1X Tris-Borate EDTA buffer adding about 40 ml. After squashing 2X Bromophenol blue solution was added to each sample as the amount of 10 ml. The samples were then centrifuged at 12600 rpm for 15 minutes at 25°C. Ten ml of supernatant was used in each slot of the gel. Seven and half percent (7.5%) gel was prepared using 7.5 ml of Acrylamide-bisacrylamide (30:0.8; Bio Basic Inc); 6.0 ml of 5X of TBE buffer; 0.15 ml of 10% ammonium persulphate (AMPS; Fulka Biochemica); 0.14 ml of TEMED and 16 ml of sterilized distilled water. The gel was run at 120V constant voltage for at least 1 hr 20 min, until the tracking dye (bromophenol blue) reached to the bottom of gel. The gel was recovered by dismantling the glass plates sinking in water very cautiously. The gel was taken in a staining tray and 30 ml of substrate mixture poured on to it for 15 min at room temperate. After 15 min, the substrate mixture was out poured and 30 ml of fast blue RR solution added to the gel and incubated at 37ºC for 25 min. Photograph of the gel was taken by a digital zoom lens camera laying the gel on a white back ground (Samsung Kenox-SHD lens, 3.2 mega pixels). Bands were scored on the stained gel from the highest to the lowest mobility and numbered accordingly. Relative mobilities were estimated towards the commonly found band Est-2 as the band for unity. Polyacrylamide gel electrophoresis (PAGE) was carried out to analyze the esterase isozyme pattern in different tissues of RJ fowl of Bangladesh. Esterases were identified in the gels by using α and β- naphthyl acetate as substrates. When only α-naphthyl acetate was hydrolyzed, the bands in the gels turned black and named as α-esterases and with only β- naphthyl acetate, the bands were red and named β-esterases. Tissue specific expression of esterase isozyme was observed in the present study. Altogether four esterase bands (Est-1, Est-2, Est-3, and Est-4) were found in RJ fowl with different relative mobilities. According to the Mendelian inheritance studies on these esterase loci, each of the bands corresponds to one allele. Data were analyzed using the general linear models procedure with the help of Statistical Package for Social Sciences (SPSS) version 10/11.5.

Source of Information: Bang. J. Anim. Sci. 2009, 38(1&2) : 15 – 21; ISSN 0003-3588

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

From the result it may be concluded that esterase gave between Red Jungle fowl and Naked Neck town has a close relationship and could be used in the field of genetics.

Knowledge Management: Journal