Halobacillus trueperi accumulates glycine betaine under condition of high osmolarity. A fragment of the glycine betaine transporter betH gene was obtained from the genome of H. trueperi with degenerate primers. Through Southern blot hybridization and inverse PCR, a 5.1 kb EcoRI fragment containing the complete betH gene was identified and subsequently sequenced. The betH gene was predicted to e...

In the pUC18-derived integration plasmid pML336 there is a 5.3-kb chromosomal DNA fragment that carries the X-prolyl dipeptidyl aminopeptidase gene (pepXP). The gene was inactivated by the insertion of an erythromycin resistance determinant into its coding sequence. Covalently closed circular DNA of pML336 was used for the electrotransformation of Lactococcus lactis. In 2% of the erythromycin-r...

SOURCE/DESCRIPTION: A 4.0 kb TaqI fragment of cosmid EFD64 isolated by a HBV-3 oligonucleotide (1) was subcloned into AccI site of pUC18. POLYMORPHISM: Mspl identifies 5 allelic VNTR polymorphism with bands between 2.6 and 4.6 kb. Rsal, TaqI, EcoRI, BamHI, Hindlll and Pvull identify the same VNTR polymorphism. FREQUENCY: With Rsal, 80 % heterozygosity were observed in 80 unrelated Caucasians. N...

SOURCE/DESCRIPTION; A 1.7 kb Mspl fragment from cosmid MCT106 isolated by an oligonucleotide (GTGTGTGTGTGTGTGTGTGT) (1) was subcloned into the AccI site of pUC18. POLYMORPHISM: PvuII identifies a three allele VNTR polymorphism with bands between 2.0 and 3.5 kb without a constant band. Mspl, TaqI, Rsal, PstI also identify the same polymorphism. FREQUENCY; 32 % hetarozygosity has been observed wi...

E. coli can be transformed to extremely high efficiencies by subjecting a mixture of cells and DNA to brief but intense electrical fields of exponential decay waveform (electroporation). We have obtained 10(9) to 10(10) transformants/micrograms with strains LE392 and DH5 alpha, and plasmids pUC18 and pBR329. The process is highly dependent on two characteristics of the electrical pulse: the ele...

Oyerexpression of the asnA gene from dseherichia coti K-12 coding for asparagine synthetase (EC 6.3.1.1) was achieved with a plasmid, pUNAd37, a derivative of pUC18, in E. cofi. The plasmid was constructed by optimizing a DNA sequence between the promoter and the ribosome binding region. The enzyme, comprising ca. 15% of the totRl soluble protei" in the E. coti cell, was readily purified to app...

The intact gtfC gene from Streptococcus mutans GS-5 was isolated in Escherichia coli in plasmid vector pUC18. The glucosyltransferase activity expressed by the gene synthesized both low-molecular-weight water-soluble glucan and insoluble glucan in a primer-independent manner. Purification of the enzyme by procedures that minimize proteolytic digestion yielded a purified preparation with a molec...

Macromolecular crowding is thought to be a significant factor driving DNA condensation in prokaryotic cells. Whereas DNA in prokaryotes is supercoiled, studies on crowding-induced DNA condensation have so far focused on linear DNA. Here we compare DNA condensation by poly(ethylene oxide) for supercoiled and linearized pUC18 plasmid DNA. It is found that supercoiling has only a limited influence...

Experiments with the supF20 mutagenesis system demonstrate that extracts from Bloom's syndrome (BS) cells are unable to use microhomology elements within the supF20 gene to restore supF function after the induction of a double-strand break (DSB). Additional experiments with the pUC18 mutagenesis system demonstrate that although the efficiency and fidelity of DSB repair by BS extracts are compar...