An Effective Electroporation Protocol for Bacillus alcalophilus TCCC11004

Author

Cheng, Kun ; Lu, Fuping ; Liang, Xiaomei

Author_Institution

Educ. Bur. Key Lab. of Ind. Microorganism, Tianjin Univ. of Sci. & Technol., Tianjin, China

fYear

2010

fDate

18-20 June 2010

Firstpage

1

Lastpage

4

Abstract

An electroporation protocol had been established for Bacillus alcalophilus TCCC11004. Parameters including growth condition of cells, electric field strength, electroporation medium, DNA concentration were evaluated in order to improve transformation efficiency. The transformation efficiency increased with electric field strength in the range of 15-20 kV/cm. The highest transformation efficiency of B. alcalophilus TCCC11004 was achieved at 20 kV/cm. The presence of osmoticums, sorbitol and mannitol, in the electroporation, growth and recovery media resulted in about 9.7 times increase in the transformation efficiency of B. alcalophilus TCCC11004, with a maximum value of 1.5×10⁴ transformants per μg of pUB110. The results demonstrated that electroporation was an effective, simple and reproducible transformation method for the B. alcalophilus TCCC11004 compared with other methods.

Keywords

DNA; bioelectric phenomena; cellular biophysics; microorganisms; molecular biophysics; protocols; Bacillus alcalophilus; DNA; TCCC11004; cell growth; electric field strength; electroporation protocol; mannitol; osmoticums; recovery media; sorbitol; transformation efficiency; Biomedical engineering; Biomembranes; Cells (biology); DNA; Fungi; Genetics; Laboratories; Microorganisms; Protocols; Purification;

fLanguage

English

Publisher

ieee

Conference_Titel

Bioinformatics and Biomedical Engineering (iCBBE), 2010 4th International Conference on

Conference_Location

Chengdu

ISSN

2151-7614

Print_ISBN

978-1-4244-4712-1

Electronic_ISBN

2151-7614

Type

conf

DOI

10.1109/ICBBE.2010.5515931

Filename

5515931