Plant Biotechnology Laboratory

  • 鄭靜明-1Plant Biotechnology Laboratory
  • Name: Ching-Ming Cheng (鄭靜明)
  •  Title: Associate professor
  •  Highest Degree: Institute of Life Science, National Tsing-Hua University, Taiwan, Ph. D.
  •  Research expertise: Plant molecular breeding, Biotechnology
  •  E-mail:lschingming@mail.tcu.edu.tw
  •  Office/ Laboratory Site: D803/D811
  •  Office/ Laboratory Tel.: +886-3-8565301 #2614/2615
  •  Research Interests:
    (1)A yeast functional expression system for positive selection of plant triterpene cyclyase
       Genetic engineering of plant secondary metabolites in Saccharomyces cerevisiae could be used to produce large amounts of desired metabolites that are either difficult to be extracted from the natural sources, or too expensive and complex to be produced by chemical synthesis. As the first step towards the production of usable pharmaceuticals in yeast, we have constructed a yeast positive selection system for direct isolation of novel terpenoid cyclyase from plants, especially from traditional Chinese herbal plants without detailed genetic or molecular information. A diploid yeast was mutated by homologous recombination to replace one copy of the erg7 gene that encodes the yeast lanosterol synthase responsible for the cyclization of 2,3-oxidosqualene. The heterozygotic mutants were selected and transformed with a recombinant copy of erg 7 using the URA3 containing centromeric plasmid pRS416-erg7. The transformants were sporulated to obtain haploid yeasts transformed with plant cDNA libraries. The URA3 on plasmid pRS416-erg7 will arrested the growth of the yeast transformants platting with 5-fluoroorotic acid (5-FOA). Only if the haploid yeasts exclude the plasmid pRS416-erg7and a plant cDNA that complements the defect of 2,3-oxidosqualene cyclization will the yeast cells get survivor. In this way, we could screen the cDNA libraries of Chinese herbal plants to isolate novel plant terpenoid synthase in a one-step positive screening network.未命名

Fig.1  Construction of the yeast functional expression system for positive selection of plant triterpene cyclyase

(2) The primary immunity determinant in modulating the lysogenic immunity of the filamentous bacteriophage cf.
To understand the immunity determinants of bacteriophage cf, we applied DNA shuffling to generate cf immunity mutants. An immunity mutant cf-ls (m-3-IG) was obtained with an infection efficiency of 106-109 fold greater than the wild type cf. Sequence comparison revealed one base increase and eight single nucleotide polymorphisms (SNPs). The nine mutations was sub-cloned to construct the cf-ls-P and cf-ls-R phages carrying the four mutations on the hypothetical PT repressor and the other five mutations upstream of the PT coding region, respectively. Phage cf-ls-P expressed an unexpected low superinfection efficiency as the wild type cf and phage cf-ls-R has completely lost its superinfection ability to the cf-lysogen. This has suggested the PT inhibitor protein and the non-coding upstream region which mediates an RNA-RNA interaction, are both involved in the regulatory mechanism of cf immunity. The result is accordance with our previous study by Cheng et al. (1999).

鄭靜明研究圖2
Fig.2  Plaque formation of cf-ls-P and cf-ls-R on XW47 and Xc G51.

  • Recent Research Grants:

NO.

associated agency

Research Grant

period

1

Tzu Chi University

Construction of a yeast functional expression system for positive screening of plant terpenoid cyclase

2012/10/1 ~

2013/9/30

2

Tzu Chi Foundation

PCR profiling of functional genes from Chinese herbal plants

2009/8/1 ~

2010/7/31

3

Tzu Chi Foundation

Isolation and characterization of ribosome-inactivating proteins from Momordica charantia

2008/8/1 ~

2009/7/31

 

  •  Research Papers:
  1. Cheng, Ching-Ming, Pelle Stolt. 2014. Basic and Applied Research on Boehmeria (Ramie) Utilising CAPS Marker Technology. Cleaved Amplified Polymorphic Sequences (CAPS) Markers in Plant Biology 11, 167-181.
  2. CI Li, SJ Chiou, TS Tong, CY Lee, LT Lee and CM Cheng. 2010. Development and validation of molecular markers for characterization of Boehmeria nivea var. nivea and Boehmeria nivea var. tenacissima. Chinese Medicine 2010, 5:40
  3. Lahmy, S., J. Guilleminot, C. M. Cheng, N. Bechtold,†, S. Albert, G. Pelletier, M. Delseny and M. Devic. 2004. DOMINO1, a member of a small plant-specific gene family encodes a protein essential for nuclear and nucleolar functions. The Plant Jounal 39, 809-820.
  4. Cheng, C. M., A. Palloix and V. Lefebvre. 2002. Isolation, mapping and characterization of allelic polymorphism of Chi-PM1, a class III chitinase of Capsicum annuum L. Plant Science 163, 481-489.
  5. Cheng, C. M., H. J. Wang, H. J. Bau and T. T. Kuo. 1999. The primary immunity determinant in modulating the lysogenic immunity of the filamentous bacteriophage cf. J. Mol. Biol. 287, 867-876.
  6. Wang, H. J., C. M. Cheng, C. N. Wang and T. T. Kuo. 1999. Transcription of the genome of the filamentous bacteriophage cf from both the plus and minus DNA strands. Virology 256, 228-232.
  7. Cheng, C. M., J. Tu, C. C. Yang and T. T. Kuo. 1996. Rifampicin an inhibitor of Xp12-specific protein phosphorylation in Xanthomonas oryzae pv. oryzae. FEMS Micro. Let. 143, 141-149.
  8. Chen, W. P., C. M. Cheng, A. H-J Wang, and T. T. Kuo. 1996. Bacteriophage cf single-stranded DNA binding protein complex purification, characterization and gene localization. Biochim. Biophy. Acta. 1309, 147-155.
  9. Huang, H. J., S. H. Lin, B. C. Yang, C. M. Cheng, C. C. Yang, and T. T. Kuo. 1995. Rapid inhibition of protein histidine phosphorylation by UV-irradiation in Xanthomonas oryzae pv. oryzae.  FEMS Micro. Let. 134, 189-194.
  10. Kuo, J. L., H. J. Huang, C. M. Cheng, L. J. Chen, B. L. Huang, L. C. Huang and T. T. Kuo. 1995. Rejuvenation in vitro: Modulation of protein phosphorylation in Sequoia sempervirens. J. Plant Physiol. 146, 333-336.
  11. Cheng, C. M., J. Tu, C. C. Yang and T. T. Kuo. 1994. Specific protein phosphorylation induced in Xanthomonas campestris pv. oryzae by bacteriophage Xp12. Arch.  Microbiol. 161, 281-285.
  12. Wu, F. S., C. M. Cheng, and T. T. Kuo. 1992. Effects of Ca++ chelator,  Ca++ ionophore, and hear shock pretreatment on in vitro protein phosphorylation of rice suspension culture cells. Bot. Bull. Academia Sinica 33, 151-159.
  13. Cheng, C. M., C. K. Chow, N. T. Hu, and T. T. Kuo. 1991. Characterization of the pp70 protein phosphorylation in the extract of rice young panicle. Biochem. Biophys. Res. Commun. 175, 467-472.
  14. Cheng, C. M., K. H. Tsai, and N. T. Hu. 1989. Protein kinase activity in divers tissues of two isogenic lines of rice. Rice Genet. Newsletter 6, 137-138.
  15. Chou C. H., S. J. Chang, C. M. Cheng, Y. C. Wang, F. H. Hsu and W. H. Den. 1989. The selective allelopathic interaction of a pasture-forest intercropping in Taiwan. II. Interaction between kikuyu grass and three hardwood plants.  Plant and Soil 116, 207-215.
  16. Cheng, C. M., N. T. Hu, and K. H. Tsai. 1987. Effects of genes Ef-1 on free amino acid content in the leaves of Taichung 65.  Rice Genet.  Newsletter 4, 96-98.

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