RT-PCR amplification of a Rhizopus oryzae lactate dehydrogenase gene fragment

Date

2001-02-01

Author

Hakki, EE
Akkaya, Mahinur

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No amino acid or DNA sequence information in sequence databases was found for a fungal lactate dehydrogenase (LDH) isozyme. Highly conserved regions in the lactate dehydrogenase enzymes of all taxonomies are found to be beta alpha beta nucleotide binding and substrate binding sites, also catalysis/active site. The conserved regions were selected as PCR primer target regions. The degenerate primers were designed according to the codon usage, determined by analyzing a number of different genes of Rhizopus species. A fragment of the gene (ldh), coding for similar to 72% of the lactate dehydrogenase enzyme from Rhizopus oryzae, was amplified using degenerate primers by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). The size of the amplified fragment containing beta alpha beta nucleotide binding site, substrate binding site and catalysis/active site is found to be about 700 bp. The reported degenerate PCR primers and the amplification conditions may lead to the cloning of the lactate dehydrogenase gene of R. oryzae, which is an important organism due to its utilization in lactic acid and enzyme productions in industrial scales. (C) 2001 Elsevier Science Inc. All rights reserved.

Subject Keywords

Biotechnology, Applied Microbiology and Biotechnology, Biochemistry

URI

https://hdl.handle.net/11511/56582

Journal

ENZYME AND MICROBIAL TECHNOLOGY

DOI

https://doi.org/10.1016/s0141-0229(00)00319-7

Collections

Graduate School of Natural and Applied Sciences, Article

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Citation Formats

E. Hakki and M. Akkaya, “RT-PCR amplification of a Rhizopus oryzae lactate dehydrogenase gene fragment,” ENZYME AND MICROBIAL TECHNOLOGY, pp. 259–264, 2001, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/56582.