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Expression of the Streptomyces coelicolor (Sc) valine (branched-chain amino acid) dehydrogenase-encoding gene (vdh) is regulated by valine, glucose and NH +4 at the transcriptional level. The results of assays for the level of accumulatedvdh mRNA in the Sc J802 strain by primer extension experiments and for the level of catechol dioxygenase (XylE) activity in Sc J802 (vdh xylE) transformants...
Sequencing of ORF10 (gene cmcH) of the Nocardia lactamdurans cephamycin gene cluster proved that it encodes a protein with a deduced molecular mass of 57 149 Da. This protein showed significant similarity to the putative O-carbamoyltransferases (O-Cases) encoded by the nodU genes of Rhizobium fredii and Bradyrhizobium japonicum, involved in the synthesis of nodulation factors. The carbamoyl-phosphate...
The nucleotide sequence of the rpoD gene (encoding the primary sigma-70 (σ 70 ) subunit of RNA polymerase) fromEscherichia coli C was determined. This gene differs from that of E. coli K-12 by a 30-bp deletion and five single-bp substitutions, resulting in a σ 70 subunit with three amino acid (aa) changes and a deletion of ten acidic aa.
The gene (TEF1) encoding translation elongation factor 1α from the dimorphic fungus Aureobasidium pullulans (Ap) strain R106 has been cloned using Candida albicans TEF1 as a heterologous hybridization probe. Electrophoretic molecular karyotype/hybridization analysis of Ap revealed eight chromosomal bands and suggested that TEF1 resides on chromosome VI. Comparison of the genomic DNA sequence with...
Recently, we described pKIL18 positive-selection cloning vectors which rely on the inactivation of the cytotoxicccdB gene [Bernard et al., Gene 148 (1994) 71-74]. They are new tools for simplifying gene cloning/sequencing procedures, as only recombinant DNA allow the formation of viable colonies. To enhance positive-selection capabilities, putative translational reinitiation codons located within...
The phosphoglycerate kinase (PGK)-encoding genes of Trypanosoma brucei are transcribed in a polycistronic fashion, but the mRNAs encoding the three PGK isozymes show differing developmental regulation. We demonstrate here that the 3'-untranslated regions of the major cytoplasmic and glycosomal PGK isozymes are capable of conferring the anticipated types of regulation on a transfected reporter gene.
NAD-dependent formate dehydrogenase (FDH) was isolated from Candida methylica (Cm) grown on 0.5% methanol. Its N-terminal amino acid (aa) sequence was determined, as was that of a commercial FDH from Candida boidinii. Degenerate oligodeoxyribonucleotides were made to the 5' region of the fdh gene from Cm using this information and to the 3' region using C-terminal aa sequence data from the methylotropic...
Expression vectors are described that make use of a plasmid-encoded nisA cassette encoding the peptide component of nisin. Specific mutations can readily be incorporated throughout the coding region of nisA. Using this vector in anisA mutant host, three variant nisins, with dehydroalanine (Dha) residues changed to Ala residues, were generated.
The TATA box, located upstream at about nt -30, and the proximal sequence element, located at about nt -60, are both essential and sufficient for basal level transcription of the Xenopus laevis (Xl) selenocysteine (Sec) tRNA [Ser]Sec gene as demonstrated by its microinjection into Xl oocytes. Point mutations within either of these regions abolish transcription,...
A series of cloning vectors has been constructed from pACYC177 and pACYC184. These vectors contain a MCS, the lacZα reporter gene, and P15A ori. Plasmids pMBL18, pMBL19 and pMBLcos express the bla gene encoding ampicillin resistance, while pMCL200 and pMCL210 contain the cat gene encoding chloramphenicol resistance. In addition, the cosmid pMBLcos was constructed with the P15A ori. These vectors...
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