Background and Aims: Since the produced recombinant proteins by molecular genetics techniques commonly have some limitations in the application, chimeric protein are introduced. Chimeric proteins have found widespread application for the study of protein folding, structure stability, function and immunogenisity. Methods: According to the known Immunomodulatory effect and structure of HSP70 mole...

We present a fast, reliable and inexpensive restriction-free cloning method for seamless DNA insertion into any plasmid without sequence limitation. Exponential megapriming PCR (EMP) cloning requires two consecutive PCR steps and can be carried out in one day. We show that EMP cloning has a higher efficiency than restriction-free (RF) cloning, especially for long inserts above 2.5 kb. EMP furth...

A rapid DNA cloning system is a research interest of many scientists. TA cloning is one of the methods used for the cloning of PCR-amplified DNA molecules. The TA cloning method is a convenient and labor-saving replacement to traditional, restriction enzyme-mediated cloning strategies. A T-vector called pBlueskript ΙΙ SK-1 with the lethal gene ccdB was designed to construct a positive selection...

A broad-host-range cloning vector, pUI81, was constructed in vitro from plasmids RSF1010 and pSL25 (a pBR322 derivative) and used to assay for transformation in Rhodopseudomonas sphaeroides. Washing cells with 500 mM Tris was an effective means of inducing competence for DNA uptake. Transformation frequencies as high as 10(-5) (transformants per viable cell) have been achieved by incubating Tri...

Background and Aims: DNA cloning, sub-cloning and site directed mutagenesis are the most common strategies in nearly all projects of recombinant protein production. The classical method of restriction site cloning is unsatisfactory due to the need for supply of restriction enzymes and the inefficiency of the digestion reaction. Many new methods, including recombinatorial cloning and ligation in...

The CAGE/GEM(TM) software toolkit for genetic engineering is briefly described. The system functionally uses color graphics and is menu driven. It integrates genetics and features information ("Overlays") with information based on sequence analysis ("Representations"). The system is structured around CAD (Computer Aided Design) principles. The CAGE (Computer Aided Genetic Engineering) aspects o...

the identification of a large number of antigens with potential for development of new tuberculosis vaccine has been accomplished in recent years. this study was designed for cloning and expression of esat-6 as a potent antigen of mycobacterium tuberculosis. selected gene (rv3875) was amplified by pcr and product was ligated into expressing plasmid vector pqe30 and recombinant pqe30-es plasmid ...

Richard Parker-Manuel, PhD, is a Group Leader for Plasmid Engineering and Production at OXGENE, A WuXi Advanced Therapies Company. He has twenty years’ experience in molecular biology with nearly seven years the cell gene therapy space. oversees design engineering of adeno associated virus lentivirus vector plasmids variety clients. also involved several R&D initiatives aim making OXGENE’s ...

A novel plasmid vector that is able to replicate both in Escherichia coli and in Streptococcus sanguis is described. This 9.2-kb plasmid, designated pVA856, carries Cmr, Tcr, and Emr determinants that are expressed in E. coli. Only the Emr determinant is expressed in S. sanguis. Both the Cmr and the Tcr of pVA856 may be insertionally inactivated. This plasmid affords several different cleavage-...