Heterologous gene expression driven by carbonic anhydrase gene promoter inDunaliella salina

Chai Yurong; Lu Yumin; Wang Tianyun; Hou Weihong; Xue Lexun

doi:10.1007/BF02842849

Your Location：

Home >

Browse articles >

Heterologous gene expression driven by carbonic anhydrase gene promoter inDunaliella salina

Updated：2023-05-16

- Heterologous gene expression driven by carbonic anhydrase gene promoter inDunaliella salina
- Journal of Oceanology and Limnology Vol. 24, Issue 4, Pages: 352-357(2006)
- Affiliations：
  
  Medical College Zhengzhou University
- Author bio：
- Funds：
- DOI：10.1007/BF02842849
  CLC：
- Online First：31 January 2013，
  
  Published：2006
- Accepted：
Scan QR Code
Yurong, C., Yumin, L., Tianyun, W. et al. Heterologous gene expression driven by carbonic anhydrase gene promoter inDunaliella salina, ., Chin. J. Ocean. Limnol. 24, 352–357 (2006). https://doi.org/10.1007/BF02842849
DOI：

Yurong, C., Yumin, L., Tianyun, W. et al. Heterologous gene expression driven by carbonic anhydrase gene promoter inDunaliella salina, ., Chin. J. Ocean. Limnol. 24, 352–357 (2006). https://doi.org/10.1007/BF02842849 DOI：

摘要

Abstract

Dunaliella salina

a halotolerant unicellular green alga without a rigid cell wall

can live in salinities ranging from 0.05 to 5 mol/L NaCl. These features ofD. salina make it an ideal host for the production of antibodies

oral vaccine

and commercially valuable polypeptides. To produce high level of heterologous proteins fromD. salina

highly efficient promoters are required to drive expression of target genes under controlled condition. In the present study

we cloned a 5′ franking region of 1.4 kb from the carbonic anhydrase (CAH) gene ofD. salina by genomic walking and PCR. The fragment was ligated to the pMD18-T vector and characterized. Sequence analysis indicated that this region contained conserved motifs

including a TATA- like box and CAAT-box. Tandem (GT)n repeats that had a potential role of transcriptional control

were also found in this region. The transcription start site (TSS) of theCAH gene was determined by 5′ RACE and nested PCR method. Transformation assays showed that the 1.4 kb fragment was able to drive expression of the selectable bar (bialaphos resistance) gene when the fusion was transformed intoD. salina by biolistics. Northern blotting hybridizations showed that the bar transcript was most abundant in cells grown in 2 mol/L NaCl

and less abundant in 0.5 mol/L NaCl

indicating that expression of the bar gene was induced at high salinity. These results suggest the potential use of theCAH gene promoter to induce the expression of heterologous genes inD. salina under varied salt condition.

关键词

Keywords

references

Apt, K. E., P. G. Kroth-Pancic and A. R. Grossman, 1996. Stable nuclear transformation of the diatomPhaeodactylum triconutum.Mol. Gen. Genet. 252: 572–579.

Avron, M. and A. Ben-Amot, 1992.Dunaliella: Physiology, Biochemistry and Biotechnogy. CRV Press, Florida, 150.

Cerutti, H., A.M. Johnson, N. Gillham and J.E. Boynton, 1997. A eubacterial gene conferring spectinomycin resistance onChlamydomonas reinhardtii: integration into the nuclear genome and gene expression.Genetics.145: 97–110.

Chai, Y.R., T.Y. Wang and L.X. Xue, 2004. Recent progress in a new bioreactor:Dunaliella salina.China Biotechnol. 24(2): 30–33. (in Chinese)

D’Halluin, K., M. De Block, J. Denecke, J. Janssens, J. Leemans, A. Reynaerts and J. Botterman, 1992. The bar gene as selectable and screenable marker in plant engineering.Methods Enzymol.216: 415–426.

Fisher, M., U. Pick and A. Zamir, 1994. A Salt-Induced 60-Kilodalton Plasma Membrane Protein Plays a Potential Role in the Extreme Halotolerance of the AlgaDunaliella.Plant Physiol.106: 1359–1365.

Geng, D.G., Y. Han, Y.Q. Wang, P. Wang, L.M. Zhang, W.B. Li and Y.R. Sun, 2004. Construction of a System for the Stable Expression of Foreign Genes inDunaliella salina.Acta Botanica Sinica.46(3): 342–346

Hallmann, A. and A. Rappel, 1999. Genetic engineering of the multicellular green alga Volvox: a modified and multiplied bacterial antibiotic resistance gene as a dominant selectable marker.Plant J.17: 99–109.

Liu, X. and M. A. Gorovsky, 1993. Mapping the 5′ and 3′ ends ofTetrahymena thermophila mRNAs using RNA ligase mediated amplification of cDNA ends (RLM-RACE).Nucl. Acid. Res. 21: 4954–4960

Liu, X. and M. A. Gorovsky, 1996. Cloning and characterization of the major histone H2A genes completes the cloning and sequencing of known histone genes ofTetrahymena thermophila.Nucleic Acids Res.16: 3023–3030.

Li, J., H. W. Ren, J.H. Huang, M.M. Yu and B. G. Ru, 2003. Transient expression of reporter gene EGFP in transformedDunaliella salina, Chinese J. Biochem Mol. Biol. 19(3): 338–342. (in Chinese)

Patnaik, D. and P. Khurana, 2003. Genetic transformation of Indian bread (T. aestivum) and pasta (T. durum) wheat by particle bombardment of mature embryo-derived calli.BMC Plant Biol. 3(1): 1521–1532.

Sambrook, J., E. F. Fritsch and T. Maniatis, 1989. Molecular Cloning: a Laboratory Manual, 2nd Cold Spring Harbor, Cold Spring Harbor Laboratory Press, NY.

Schiedlmeier, B., R. Schmitt, W. Muller, M. M. Kirk, H. Gruber, W. Mages and D.L. Kirk, 1994. Nuclear transformation ofVolvox carteri.Proc. Natl. Acad. Sci. U.S.A. 91: 5080–5084.

Shimogawara, K., S. Fujiwarab, A.R. Grossman and H. Usudaa, 1998. High-Efficiency Transformation ofChlamydomonas reinhardtii by Electroporation.Genetics.148: 1821–1828.

Stacey, J. and P.G. Isaac, 1994. Isolation of DNA from plants.Methods Mol. Biol. 28: 9–15.

Stevens, D. R., J. D. Rochaix and S. Purton, 1996. The bacterial phleomycin resistance geneble as a dominant selectable marker inChlamydomonas.Mol. Gen. Genet. 251: 23–30.

Zhang, G. X., P. R. Xu, H. M. Zheng, J. M. Wang and L. X. Xue, 2002. The resistance ofDunaliella salina to the herbicide phosphinothricin.Life Sci. Res. 6(1): 92–94.

Views

Downloads

CSCD

Alert me when the article has been cited

Submit

Tools

Publicity Resources

Hydrochemical characteristics and the ecological effect of algal carbonic anhydrase in carbon cycle in the Taiyuan section of Fenhe River

Functional analysis of the promoter of the dmrt1 gene in Chinese tongue sole, Cynoglossus semilaevis

Effect of nitrate and phosphate on accumulation of β-carotene isomers inDunaliella salina

Effects of Mg2+, NaCl, citric acid, and other factors on synthesis and accumulation of β-carotene inDunaliella salina

Numerical Studies on Effects of V-Shaped Leading Edge on Incident Shock Wave Boundary Layer Interaction

Related Author

Jia FENG

Shulian XIE

Xin LI

Jing YANG

Yingming YANG

Qian WANG

Zhongkai CUI

Hua GUO

Related Institution

Shanxi Key Laboratory of Yuncheng Salt Lake Ecological Protection and Resource Utilization, College of Life Sciences, Yuncheng University

School of Life Science, Shanxi University

Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences

Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology

Institute of Oceanology

Address：Editorial Office of JOL, 88 Haijun Rd. Qingdao, 266000, China
Tel：+86-532-82898754 Email：jol@qdio.ac.cn
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰