The cAMP receptor protein (CRP, also known as CAP) functions as a transcriptional activator at more than 100 different E. coli operons (3). CRP is arguably one of the most well-understood transcriptional activators. At simple CRP-dependent promoters, where CRP is the only activator, transcription activation by CRP involves direct contact with RNA polymerase at two distinct sites. These contacts...

reteplase is a segment of tissue plasminogen activator used for the removal of thrombi in blood vessels. in the present study the cloned reteplase gene was used for its expression in competent e. coli. the  recombinant plasmid, pet15b/reteplase (rpet-bl21), was transformed into competent e.coli strain bl21 (de3) cells. overnight culture of the transformed bacteria was induced by the addition of...

alkaline phosphatase is an enzyme with widespread use in research and industry such as protein labeling, dephosphorylation of nucleic acids, and enzyme based biosensors. in the present study, alkaline phosphatase gene was inserted into the pet15b vector. the recombinant dna was then expressed using iptg as an inducer. the expression of this enzyme was optimized by changing expression conditions...

BACKGROUND Plasminogen activator inhibitor type-2 (PAI-2, SERPINB2) is an irreversible, specific inhibitor of the urokinase plasminogen activator (uPA). Since overexpression of uPA at the surface of cancer cells is linked to malignancy, targeting of uPA by exogenous recombinant PAI-2 has been proposed as the basis of potential cancer therapies. To this end, reproducible yields of high purity pr...

The complete decipherment of the functions and interactions of the elements of the riboflavin biosynthesis operon (rib operon) of Bacillus subtilis are necessary for the development of superproducers of this important vitamin. The function of its terminal ribT gene has not been established to date. In this work, a search for homologs of the hypothetical amino acid sequence of the gene product t...

The VPl gene of enterovirus 71 (EV71) was synthesized, construct a recombinant plasmid pET15b/VP1 and expressed in E. coli BL21. The recombinant VP1 protein could specifically react with EV71-infected patient sera without the cross-reaction with serum antibodies of coxsackievirus A16 (CA16), A4, A5, B3 and B5 as well as echovirus 6. In acute and convalescent phases, IgM and IgG antibodies of 18...

Major immune-dominant proteins of Leptospira interrogans Icterohaemorrhagiae were cloned, expressed in prokaryotic system was purified and confirmed by western blot. The protective efficacy of purified recombinant proteins was assayed synergistically with adjuvant in hamster models. Three combination were assessed for homologous protection (i.e, rLipL41, rOmpL1, rHapI and rSphH; rLipL41, rOmpL1...

The ywtD gene, which codes for an enzyme that degrades -polyglutamic acid (PGA), was cloned from Bacillus subtilis IFO16449. The gene is located immediately downstream of ywsC and ywtABC, a PGA operon involved in PGA biosynthesis, and it showed partial similarity to genes coding for DL-endopeptidase, a peptidoglycan-degrading enzyme. The ywtD gene, from which signal sequence is excised, was ins...

The ywtD gene, which codes for an enzyme that degrades gamma-polyglutamic acid (PGA), was cloned from Bacillus subtilis IFO16449. The gene is located immediately downstream of ywsC and ywtABC, a PGA operon involved in PGA biosynthesis, and it showed partial similarity to genes coding for DL-endopeptidase, a peptidoglycan-degrading enzyme. The ywtD gene, from which signal sequence is excised, wa...

With the help of sepiolite, a unique method for transforming DNA into bacteria, based on the Yoshida effect, has been developed recently. However, we confronted many problems when this newest method was tried. Only a few transformants could be obtained even when 100 ng of plasmid pET15b was used, and a successful result seemed difficult to repeat. To address this problem, we optimized the opera...