Cloning and Expression of the Haemagglutinin-neuraminidase Gene of Newcastle Disease Virus Strain Af2240 Lnto Eschericia Coli

A 1815 bp sequence of the open reading frame (ORF) of the haemagglutinin-neuraminidase (HN) gene of velogenic-viscerotropic Newcastle disease virus (NDV) strain AF2240 was amplified reaction (PCR) using primers SI (5'CACCAATAGCAGACTCCA3') and S2 (5'CCTTGGCATTGCAGAAG3'). Both p...

Full description

Saved in:

Bibliographic Details
Main Author:	Sudin, Sudani
Format:	Thesis
Language:	English English
Published:	1997
Subjects:	Newcastle disease virus Eschericia Coli
Online Access:	http://psasir.upm.edu.my/id/eprint/8630/1/FSAS_1997_15_A.pdf
Tags:	Add Tag No Tags, Be the first to tag this record!

id	my-upm-ir.8630
record_format	uketd_dc
spelling	my-upm-ir.86302012-05-08T08:55:02Z Cloning and Expression of the Haemagglutinin-neuraminidase Gene of Newcastle Disease Virus Strain Af2240 Lnto Eschericia Coli 1997 Sudin, Sudani A 1815 bp sequence of the open reading frame (ORF) of the haemagglutinin-neuraminidase (HN) gene of velogenic-viscerotropic Newcastle disease virus (NDV) strain AF2240 was amplified reaction (PCR) using primers SI (5'CACCAATAGCAGACTCCA3') and S2 (5'CCTTGGCATTGCAGAAG3'). Both primers had dUMP extensions which allowed direct cloning of the amplified product into the pAMP1 vector, which also has the complementary protruding termini.The recombinant plasmid was transformed into competent Eschericia coli strain DH5u. A total of 174 transformants were obtained and the transformation efficiency was 3.8 x 10⁴ CFU/µg. The presence of the HN specific inserts in the transformants were detected by colony hybridization, Southern blotting and finally nucleotide sequence analysis. The HN protein was shown to be expressed as an unglycosylated protein of size 63.0 kDa which is comparable to that predicted from the nucleotide sequence. Thus, this shows that the HN protein was successfully expressed from the E. coli and the protein obtained could be used for further analysis. Newcastle disease virus Eschericia Coli 1997 Thesis http://psasir.upm.edu.my/id/eprint/8630/ http://psasir.upm.edu.my/id/eprint/8630/1/FSAS_1997_15_A.pdf application/pdf en public masters Universiti Putra Malaysia Newcastle disease virus Eschericia Coli Faculty of Environmental Studies English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Newcastle disease virus Eschericia Coli
spellingShingle	Newcastle disease virus Eschericia Coli Sudin, Sudani Cloning and Expression of the Haemagglutinin-neuraminidase Gene of Newcastle Disease Virus Strain Af2240 Lnto Eschericia Coli
description	A 1815 bp sequence of the open reading frame (ORF) of the haemagglutinin-neuraminidase (HN) gene of velogenic-viscerotropic Newcastle disease virus (NDV) strain AF2240 was amplified reaction (PCR) using primers SI (5'CACCAATAGCAGACTCCA3') and S2 (5'CCTTGGCATTGCAGAAG3'). Both primers had dUMP extensions which allowed direct cloning of the amplified product into the pAMP1 vector, which also has the complementary protruding termini.The recombinant plasmid was transformed into competent Eschericia coli strain DH5u. A total of 174 transformants were obtained and the transformation efficiency was 3.8 x 10⁴ CFU/µg. The presence of the HN specific inserts in the transformants were detected by colony hybridization, Southern blotting and finally nucleotide sequence analysis. The HN protein was shown to be expressed as an unglycosylated protein of size 63.0 kDa which is comparable to that predicted from the nucleotide sequence. Thus, this shows that the HN protein was successfully expressed from the E. coli and the protein obtained could be used for further analysis.
format	Thesis
qualification_level	Master's degree
author	Sudin, Sudani
author_facet	Sudin, Sudani
author_sort	Sudin, Sudani
title	Cloning and Expression of the Haemagglutinin-neuraminidase Gene of Newcastle Disease Virus Strain Af2240 Lnto Eschericia Coli
title_short	Cloning and Expression of the Haemagglutinin-neuraminidase Gene of Newcastle Disease Virus Strain Af2240 Lnto Eschericia Coli
title_full	Cloning and Expression of the Haemagglutinin-neuraminidase Gene of Newcastle Disease Virus Strain Af2240 Lnto Eschericia Coli
title_fullStr	Cloning and Expression of the Haemagglutinin-neuraminidase Gene of Newcastle Disease Virus Strain Af2240 Lnto Eschericia Coli
title_full_unstemmed	Cloning and Expression of the Haemagglutinin-neuraminidase Gene of Newcastle Disease Virus Strain Af2240 Lnto Eschericia Coli
title_sort	cloning and expression of the haemagglutinin-neuraminidase gene of newcastle disease virus strain af2240 lnto eschericia coli
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Environmental Studies
publishDate	1997
url	http://psasir.upm.edu.my/id/eprint/8630/1/FSAS_1997_15_A.pdf
_version_	1747810843346599936

Cloning and Expression of the Haemagglutinin-neuraminidase Gene of Newcastle Disease Virus Strain Af2240 Lnto Eschericia Coli

Similar Items