Characterization of a halobacterial gene affecting bacterio-opsin gene expression.

نویسندگان

  • M Betlach
  • J Friedman
  • H W Boyer
  • F Pfeifer
چکیده

A substantial number of spontaneous bacterio-opsin mutants of Halobacterium halobium are the result of insertion elements up to 1400 bp upstream of the bacterio-opsin (bop) gene. The nucleotide sequence of 1800 bp upstream of the bop gene has been determined. There is a 1118 bp open reading frame (ORF) located within this region which is transcribed and which coincides with the distribution of insertion elements upstream of the bop gene in Bop mutants. Therefore, we propose that there is a gene (brp gene) 526 bp upstream of the bop gene. This putative gene is transcribed in the opposite direction as the bop gene and could encode a protein of 37,500 D (359 amino acids) with a codon usage similar to bacterio-opsin. The 5' terminus of the brp transcript has been determined. The brp transcript and the bop mRNA are complementary for 13 residues near their 5' termini and both transcripts start at or near the initiating codon of the gene. Both transcripts could form similar hairpin loop structures at their 5' termini which contain possible ribosomal binding sites. The DNA sequences immediately upstream of the bop and the brp genes have significant homologies and there is a short complementary sequence. The role of the brp gene in bacterio-opsin gene expression is unclear.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Characterization of insertions affecting the expression of the bacterio-opsin gene in Halobacterium halobium.

We have determined the sequence of the inverted repeats and duplicated target DNA of the halobacterial insertion elements ISH2 (520 bp), ISH23 (900 bp) and ISH24 (3000 bp) associated with bacterio-opsin (bop) mutants. ISH2 has a perfect 19 bp inverted repeat (3,5), while both ISH23 and ISH24 have imperfect inverted repeats of 29 bp and 14 bp respectively. ISH23 was shown to be highly homologous...

متن کامل

Studies on the biosynthesis of bacterio-opsin. Demonstration of the existence of protein species structurally related to bacterio-opsin.

1. The kinetics of processing newly synthesized bacterio-opsin from the non-crystalline state within the brown membrane to the crystalline state within the purple membrane was followed by pulse-chase experiments. 2. Biosynthesis of bacterio-opsin was found to be highly resistant to RNA-synthesis inhibitors like rifampicin and ethidium bromide. In the presence of ethidium bromide, only five prot...

متن کامل

Homologous bacterio-opsin-encoding gene expression via site-specific vector integration.

Homologous recombination in the archaebacterium Halobacterium halobium has been investigated and exploited for the wild-type (wt) level of expression of the bacterio-opsin-encoding gene (bop). The Haloferax volcanii-Escherichia coli shuttle vector, pWL102, was used to construct a shuttle-mutagenesis vector, pEF191, bearing bop and short flanking sequences. Transformation of a bacteriorhodopsin ...

متن کامل

Characterization of a second gene involved in bacterio-opsin gene expression in a halophilic archaebacterium.

Southern blot analysis and nucleotide sequencing of DNA from three bacterio-opsin-deficient mutants of the archaebacterium Halobacterium halobium (M86, W105, and W109) revealed that they each contain an alteration in a region 2,000 to 3,800 base pairs (bp) upstream of the bacterio-opsin gene (bop). Nucleotide sequence analysis of this region, which is also located downstream of the previously c...

متن کامل

A C-terminal truncation results in high-level expression of the functional photoreceptor sensory rhodopsin I in the archaeon Halobacterium salinarium.

Expression of the gene encoding the halobacterial photoreceptor sensory rhodopsin I (SRI), sopI, was studied by means of homologous gene targeting. A sopI- Halobacterium salinarium mutant strain was constructed by homologous replacement of sopI with a novobiocin-resistant gyrB from Haloferax Aa 2.2. Cells bearing gyrB were resistant to novobiocin, indicating that the Haloferax gene is functiona...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Nucleic acids research

دوره 12 20  شماره 

صفحات  -

تاریخ انتشار 1984