NISCAIR Online Periodicals Repository

NISCAIR ONLINE PERIODICALS REPOSITORY (NOPR)  >
NISCAIR PUBLICATIONS >
Research Journals >
Indian Journal of Biochemistry and Biophysics (IJBB) >
IJBB Vol.46 [2009] >
IJBB Vol.46(3) [June 2009] >


Title: Single-stranded megaprimer splicing through OE-PCR: Construction of full-length Aspergillus niger arginase cDNA
Authors: Jayashri, T N
Anuradha, R
Punekar, N S
Keywords: Single-stranded megaprimer
Overlap extension PCR
Splicing
Aspergillus niger
Arginase
Issue Date: Jun-2009
Publisher: CSIR
Abstract: A useful variant of PCR technique was devised to generate full-length Aspergillus niger arginase cDNA for expression. Briefly, a 450 bp amplicon was first constructed through overlap extension PCR (OE-PCR) by splicing in a 101 nucleotide long single-stranded megaprimer, facilitated by inclusion of an additional, shorter forward primer in the reaction. The amplicon was suitably cloned into pBlueScript to obtain pArg440 and the insert sequenced. The full-length arginase cDNA was subsequently assembled in pArg440 and moved into pET23a for heterologous expression. An interesting feature of this strategy was not to stoichiometrically incorporate the oligonucleotide megaprimer, but use it only as an early template. This OE-PCR strategy to utilize long single-stranded megaprimer may prove handy in terms of efficiency, yield and sequence choice.
Page(s): 266-268
ISSN: 0975-0959 (Online); 0301-1208 (Print)
Source:IJBB Vol.46(3) [June 2009]

Files in This Item:

File Description SizeFormat
IJBB 46(3) 266-268.pdf971.55 kBAdobe PDFView/Open
 Current Page Visits: 879 
Recommend this item

 

National Knowledge Resources Consortium |  NISCAIR Website |  Contact us |  Feedback

Disclaimer: NISCAIR assumes no responsibility for the statements and opinions advanced by contributors. The editorial staff in its work of examining papers received for publication is helped, in an honorary capacity, by many distinguished engineers and scientists.

CC License Except where otherwise noted, the Articles on this site are licensed under Creative Commons License: CC Attribution-Noncommercial-No Derivative Works 2.5 India

Copyright © 2012 The Council of Scientific and Industrial Research, New Delhi. All rights reserved.

Powered by DSpace Copyright © 2002-2007 MIT and Hewlett-Packard | Compliant to OAI-PMH V 2.0

Home Page Total Visits: 513366 since 06-Feb-2009  Last updated on 21-Apr-2014Webmaster: nopr@niscair.res.in