Cloning and molecular characterization of delta12-fatty acid desaturase gene from Mortierella isabellina.

ABSTRACT:

Aim

To clone delta(12)-fatty acid desaturase gene of Mortierella isabellina and to functionally characterize this gene in vitro and in vivo.

Methods

Reverse transcriptional polymerase chain reaction (RT-PCR) was used to clone the open reading frame of delta(12)-fatty acid desaturase gene (D12D) of Mortierella isabellina. Plasmids pEMICL12 and pYMICL12 were constructed with it. pEMICL12 was transformed into Escherichia coli (E.coli) strain BL21 using CaCl(2) method for expression after induction with IPTG. pTMICL12 was transformed into Saccharomyces cerevisiae strain INVSc1 using lithium acetate method for expression under the induction of galactose. Northern blotting method was used to investigate the effect of temperature on the transcriptional level of this gene in S.cerev

SUBMITTER: Li MC 

PROVIDER: S-EPMC4087868 | biostudies-literature | 2006 Jun

REPOSITORIES: biostudies-literature