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Applied and Environmental Microbiology, September 1999, p. 3888-3895, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Analysis of the Gene Family Encoding Lipases in Candida rugosa by Competitive Reverse Transcription-PCR

Guan-Chiun Lee,1 Shye-Jye Tang,2,* Kuang-Hui Sun,3 and Jei-Fu Shaw1,4,*

Institute of Marine Biotechnology, National Taiwan Ocean University, Keelung, Taiwan 20224,2 Institute of Biochemistry,1 and Department of Medical Technology3, National Yang-Ming University, Taipei, Taiwan 11211, and Institute of Botany, Academia Sinica, Taipei, Taiwan 115294

Received 8 March 1999/Accepted 6 June 1999

Synthesis of multiple extracellular lipases in Candida rugosa has been demonstrated. However, it is difficult to characterize the expression spectrum of lip genes, since the sequences of the lip multigene family are very closely related. A competitive reverse transcription-PCR assay was developed to quantify the expression of lip genes. In agreement with the protein profile, the abundance of lip mRNAs was found to be (in decreasing order) lip1, lip3, lip2, lip5, and lip4. To analyze the effects of different culture conditions, the transcript concentrations for these mRNA species were normalized relative to the values for gpd, encoding glyceraldehyde-3-phosphate dehydrogenase. In relative terms, lip1 and lip3 were highly and constitutively expressed (about 105 molecules per µg of total RNA) whereas the other inducible lip genes, especially lip4, showed significant changes in mRNA expression under different culture conditions. These results indicate that differential transcriptional control of lip genes results in multiple forms of lipase proteins.

* Corresponding authors. Mailing address for Dr. Shaw: Institute of Botany, Academia Sinica, Nankang, Taipei, Taiwan 11529. Phone: 886-2-27899590 ext. 226. Fax: 886-2-27821605. E-mail: boplshaw{at}gate.sinica.edu.tw. Mailing address for Dr. Tang: Institute of Marine Biotechnology, National Taiwan Ocean University, Keelung, Taiwan 20224. Phone: 886-2-24622192 ext. 5510. Fax: 886-2-24622320. E-mail: tsj{at}ntou-66.ntou.edu.tw.

Applied and Environmental Microbiology, September 1999, p. 3888-3895, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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