Journal of Food, Agriculture and Environment

Vol 12, Issue 2,2014
Online ISSN: 1459-0263
Print ISSN: 1459-0255

Single-point mutation as a molecular tool for preparation of recombinant vaccine against Caseous Lymphadenitis


Ihab Mohamed Moussa 1*, Ashgan Mohamed Hessain 2, 3, Hassan Abdullah Hemeg 4, Salha Abdelkareem Selim 3

Recieved Date: 2014-01-13, Accepted Date: 2014-03-28


The gene encoding the phospholipase D exotoxin (PLD) of Corynebacterium pseudotuberculosis; the causative agent of Caseous Lymphadenitis (CLA) in sheep and Oedematous Skin Disease (OSD) in buffaloes was mutagenised using PCR-site directed mutagenesis. Mutagenesis was performed to substitute Histidine codon 20 (CAC) with Tyrosine codon 20 (TAT) within the enzymatic active site of the pld gene. The wild and mutated pld genes were expressed in Escherichia coli purified from cell lysate using calmodulin affinity chromatography (stratagene). A comparative study about synergistic hemolytic activity, dermonecrosis; immunogenicity and antigencity of both recombinants PLD (rPLD) was performed. Results showed that mutated recombinant PLD (mrPLD) had lost the synergistic hemolytic activity (SH), but both had the same characteristic in relation to dermonecrosis, immunogenic and antigenic activities. The results suggest that rPLD can be recommended as highly specific antigens for early diagnosis of CLA in small ruminants in addition to form the basis for recombinant PLD vaccines.


Wild recombinant PLD, mutated recombinant PLD, dermonecrotic activity, immunogencity of rPLD, mutagenesis

Journal: Journal of Food, Agriculture and Environment
Year: 2014
Volume: 12
Issue: 2
Category: Agriculture
Pages: 626-629

Full text for Subscribers

Note to users

The requested document is freely available only to subscribers/registered users with an online subscription to the Journal of Food, Agriculture & Environment. If you have set up a personal subscription to this title please enter your user name and password. All abstracts are available for free.

Article purchasing

If you like to purchase this specific document such as article, review or this journal issue, contact us. Specify the title of the article or review, issue, number, volume and date of the publication. Software and compilation, Science & Technology, all rights reserved. Your use of this website details or service is governed by terms of use. Authors are invited to check from time to time news or information.

Purchase this Article:   20 Purchase PDF Order Reprints for 15

Share this article :