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