Insertion sequence IST3091 of Thiobacillus ferrooxidans

• Published in: Canadian journal of microbiology Vol. 43; no. 6; p. 503
• Main Authors: Chakravarty, L, Kittle, Jr, J D, Tuovinen, O H
• Format: Journal Article
• Language: English
• Published: Canada 01.06.1997
• Subjects: Amino Acid Sequence; Base Sequence; Cloning, Molecular; Conjugation, Genetic; DNA Primers - genetics; DNA Transposable Elements - genetics; DNA, Bacterial - genetics; Escherichia coli - genetics; Genes, Bacterial; Genome, Bacterial; Molecular Sequence Data; Plasmids - genetics; Polymerase Chain Reaction; Protein Biosynthesis; Repetitive Sequences, Nucleic Acid; Replication Origin - genetics; Sequence Homology, Amino Acid; Sequence Homology, Nucleic Acid; Thiobacillus - genetics
• ISSN: 0008-4166; 1480-3275
• DOI: 10.1139/m97-072

An insertion sequence, designated as IST3091, was located adjacent to the putative origin of replication region of plasmid pTFI91 of Thiobacillus ferrooxidans TFI-91. The DNA sequence of the transposase gene of IST3091 revealed similarity with that of IS30, IS1086, IS4351, and the integrase gene of SpV1-R8A2 B (a bacteriophage of Spiroplasma citri). The sequence of IST3091 is 1063 bp long with partially matched 30-bp terminal inverted repeats. Several restriction fragments of plasmid pTFI91 of T. ferrooxidans containing the IST3091 element were cloned into the vector pHSG398. The hybrid plasmids (pBTL) were transformed into Escherichia coli NK7379 containing a miniF plasmid, which was devoid of transposable elements. The transposition function of the IST3091 element was confirmed by mobilizing hybrid plasmids via conjugation from transformed E. coli NK7379 (donor) to E. coli M8820 (recipient). The presence of the transposed element in transconjugants was detected by polymerase chain reaction amplification.