Biochemistry

Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 9 No. 3, Issue of April 15, 2006
© 2006 by Pontificia Universidad Católica de Valparaíso -- Chile  
DOI: 10.2225/vol9-issue3-fulltext-16  
RESEARCH ARTICLE

Production of recombinant enzymes of wide use for research

María J. Manzur
Departamento de Bioquímica y Ciencias Biológicas
Facultad de Química, Bioquímica y Farmacia
Universidad Nacional de San Luis
Ejército de los Andes 950
San Luis, Argentina
Tel/Fax: 54 2652 422644
E-mail: mjmanzu@unsl.edu.ar

Rosana V. Muñoz
Departamento de Bioquímica y Ciencias Biológicas
Facultad de Química, Bioquímica y Farmacia
Universidad Nacional de San Luis
Ejército de los Andes 950
San Luis, Argentina
Tel/Fax: 54 2652 422644
E-mail: munoz@bio.puc.cl

 Adrián A. Lucero
Departamento de Bioquímica y Ciencias Biológicas
Facultad de Química, Bioquímica y Farmacia
Universidad Nacional de San Luis
Ejército de los Andes 950
San Luis, Argentina
Tel/Fax: 54 2652 422644
E-mail: adrian_lucerhoff@yahoo.com 

 Maximiliano Juri Ayub
Departamento de Bioquímica y Ciencias Biológicas
Facultad de Química, Bioquímica y Farmacia
Universidad Nacional de San Luis
Ejército de los Andes 950
San Luis, Argentina
Tel/Fax: 54 2652 422644
E-mail: juriayub@dna.uba.ar

 Sergio E. Alvarez
Departamento de Bioquímica y Ciencias Biológicas
Facultad de Química, Bioquímica y Farmacia
Universidad Nacional de San Luis
Ejército de los Andes 950
San Luis, Argentina
Tel/Fax: 54 2652 422644
E-mail: sealvar@unsl.edu.ar

 Gladys M. Ciuffo*
Departamento de Bioquímica y Ciencias Biológicas
Facultad de Química, Bioquímica y Farmacia
Universidad Nacional de San Luis
Ejército de los Andes 950
San Luis, Argentina
Tel/Fax: 54 2652 422644
E-mail: gciuffo@unsl.edu.ar

*Corresponding author

Financial support: Grant from the Universidad Nacional de San Luis, Argentina.

Keywords: bioactivity, protein expression, purification, recombinant enzymes.

Abbreviations:

Ang II: Angiotensin II
AT2: Angiotensin II type 2 receptor
GAPDH: Glyseraldehyde-3-phosphate dehydrogenase
MMLV: Moloney murine Leukemia Virus
PND: post-natal day
SN: supernatant


Abstract
Reprint (PDF)

For biotechnological purposes, protein expression refers to the directed synthesis of large amounts of desired proteins. The aim of the present work was to produce reverse transcriptase Moloney murine Leukaemia Virus retro-transcriptase and Taq DNA polymerase, as bioactive products. In the present paper, we report the preparation of recombinant enzymes, expressed in E. coli strains. The enzymes produced exhibited quite good activity, compared with commercial enzymes, allowing us to replace the last ones for several lab applications. We are reporting changes and modifications to standard protocols described. The standard protocols were modified, i.e. for the purification step of Taq, a temperature dependent procedure was designed. The enzymes produced were used in different applications, such as PCR, RT-PCR, PCR Multiplex and RAPDs molecular markers.

 
Supported by UNESCO / MIRCEN network 
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