[Cloning the uridine phosphorylase (udp) gene of Escherichia coli and its expression in recombinant plasmids].
On the basis of Escherichia coli DNA and vectors pBR322, pUC19, hybrid plasmids restoring Udp+ phenotype in the E. coli deletion (delta udp) mutant have been obtained. The udp gene is carried by a 8 kb PstI fragment (on the pUD2) and by a smaller 2.87 kb PstI-SalGI fragment from the PstI fragment (pUD7). The uridine phosphorylase level was 30 times higher in the cells containing hybrid plasmid as compared to the strain with chromosomal location of the udp gene. On the other hand, the measurements of uridine phosphorylase activity in the cytR- and cya- background indicate that expression of the cloned udp gene escapes partially negative control of the CytR repressor and positive control of cAMP--CRP complex. These data suggest that the 2.87 kb PstI--SalGI-fragment contains the intact udp gene which is transcribed from its own promoter. Increase in the activity of beta-galactosidase encoded by udp-lacZ fusion has been observed in the presence of pUD2 or pUD7, which was suggested to be the consequence of titration of CytR repressor molecules in the operator region of the cloned udp.
Brikun
I A
IA
Mironov
A S
AS
Sukhodolets
V V
VV
Khurges
E M
EM
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D007763:Lac Operon; D010430:Pentosyltransferases / Q000235:genetics*; D010957:Plasmids; D011995:Recombination, Genetic; D015183:Restriction Mapping; D014543:Uridine Phosphorylase / Q000235:genetics*; D001616:beta-Galactosidase / Q000378:metabolism
D010430:Pentosyltransferases; D014543:Uridine Phosphorylase; D001616:beta-Galactosidase
Klonirovanie gena uridinfosforilazy (UDP) Escherichia coli i ego ékspressiia v sostave rekombinantnykh plazmid.