NUCLEOTIDE-SEQUENCE AND OVEREXPRESSION OF THE TELLURITE-RESISTANCE DETERMINANT FROM THE INCHII PLASMID PHH1508A

作者：WALTER EG; WEINER JH; TAYLOR DE

来源：Gene, 1991, 101(1): 1-7.

DOI：10.1016/0378-1119(91)90217-Y

• RECOMBINANT DNA
• TRANSPOSON INSERTION MUTAGENESIS
• FUSION PROTEINS
• METAL RESISTANCE
• TRANSCRIPTIONAL ANALYSIS
• RNA PRIMER EXTENSION
• PROTEIN OVERPRODUCTION

The transcription and translation of the tellurite-resistance (Te(R)) genes of the HII incompatibility group plasmid, pHH1508a, were studied. The nucleotide (nt) sequence of the Te(R) region was determined and two possible open reading frames, tehA and tehB, were identified. The direction of transcription and translation of these genes was confirmed through the preparation of lacZ and phoA (encoding alkaline phosphatase) fusions. The transcription start point was identified in the sequence using RNA primer extension. The tehA gene codes for a 36-kDa polypeptide which is highly hydrophobic. The TehA protein appears to be located in the inner membrane of the bacterial cell since tehA fusions with both phoA and lacZ were obtained and expressed. The tehB gene codes for a 23-kDa polypeptide which appears to be relatively hydrophilic and is probably located in the cytoplasm. Both proteins were overproduced using a T7 RNA polymerase/promoter system. No nt or amino acid sequence homology could be found between this Te(R) determinant and the Te(R) genes from the IncHI-2 plasmid, pMER610, and the IncP-alpha-plasmid, RK2.

• 出版日期1991-5-15