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EXPRESSION OF THE DETOXIFYING GENE B1 IN ES‐CHERICHIA COLI AND SYNECHOCOCCUS 1
Author(s) -
Yanchun YAN,
Chuanlin QIAO,
Hongyu SHANG
Publication year - 2000
Publication title -
insect science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 45
eISSN - 1744-7917
pISSN - 1672-9609
DOI - 10.1111/j.1744-7917.2000.tb00355.x
Subject(s) - biology , shuttle vector , microbiology and biotechnology , plasmid , southern blot , blot , complementary dna , esterase , expression vector , escherichia coli , gene , recombinant dna , clone (java method) , kanamycin , northern blot , vector (molecular biology) , enzyme , biochemistry
We inserted the mosquito esterase B1 gene into the expression vector pRL‐439, which possesses the strong promoter PpsbA. The recombinant plasmid pRL‐Bl was used to transform E. coli HBlOl, and the positive clones were screened on LB medium plate containing 100 mg/mL ampicillin. The results of dot blotting and Southern hybridization demonstrated that these positive clones were transformed bacteria. Western blotting indicated that esterase B1 gene had been successfully expressed under the control of the PpsbA promoter in E. coli . A shuttle verruction‐B1 (pDGBl) was constructed by inserting B1‐cDNA from pRL‐Bl into polycloning site of plasmid pDc8. PDGBl was transferred into Synechoccus sp. PCC7942 through triparental conjugal transfer. Transformed single Synechococcus sp. PCC 7942 clone was obtained by neomycin screening, and large‐scale culture in liquid medium was carried out. Results of Southern blotting proved that pWB1 was transferred into Synechococcus sp. PCC 7942.