background: bone morphogenetic protein 2 (bmp- 2) belongs to the tgf-β superfamily of proteins and plays an important role in the development of bone and cartilage. bmp- 2 is also associated with maintenance and repair of damaged bone. recombinant human bone morphogenetic protein 2 (rhbmp- 2) is now produced by genetic engineering techniques and used in treatment of thin bone fractures in the j...

Since the construction of the first generation of general cloning vectors in the early 1970s, the number of plasmids created has increased to an almost countless number. Thus, a critical decision facing today’s investigator is that of which plasmid to use in a particular project? Despite the bewildering choice of commercial and other available vectors, the choice of which cloning vector to use ...

We previously expressed VbhTA constructs for lysate AMPylation assays from the pRSFDuet-1 backbone, a plasmid with two multiple cloning sites that each enable the expression of constructs from a PT7 promoter. Our constructs encoded either the VbhT toxin alone (with an N-terminal hexahistidine (His6-)tag) to be expressed from the first multiple cloning site or had additionally the VbhA antitoxin...

objective: in this project, our aim was to construct a novel expressing vector harboring a new sequence from overlapping region of ns3 gene of hcv from infected iranian patient. materials and methods: the partial ns3 (pns3) gene was amplified by nested-rt-pcr method using sera of hcv infected patients harboring genotype 1a. after purification and cloning the pns3 into ta-cloning vector, the be...

Two unique isolates of Borrelia burgdorferi, differing in plasmid content and outer surface protein C expression, were cultured on sequential captures of a single free-living Peromyscus leucopus mouse and were examined for differences in transmissibility. Both isolates were transmissible from inoculated C.B-17 mice to larval Ixodes scapularis ticks and, subsequently, from infected nymphal ticks...

This report describes the first functional analysis of a bacteriocin immunity gene from Propionibacterium freudenreichii and its use as a selection marker for food-grade cloning. Cloning of the pcfI gene (previously orf5 [located as part of the pcfABC propionicin F operon]) rendered the sensitive host 1,000-fold more tolerant to the propionicin F bacteriocin. The physiochemical properties of th...

Methodology: We tissue cultured the PVX positive potato plants for viral RNA extraction. Total RNA was converted to cDNA for priming CP gene in PCR for amplification. To get the complete sequence of gene, we cloned CP gene into pTZ57R/T cloning vector. Upon double digestion of recombinant plasmid with EcoRI and HindIII restriction enzymes, 710 bp fragment was obtained which confirmed cloning. R...

We describe a simple PCR-based method for the isolation of genomic DNA that lies adjacent to a known DNA sequence. The method is based on the directional cloning of digested genomic DNA into the multiple cloning site of a pUC-based plasmid to generate a limited genomic library. The library is plated onto a number of selective LA plates which are incubated overnight, and recombinant plasmid DNA ...

A method for positional cloning of the Bacillus subtilis genome was developed. The method requires a set of two small DNA fragments that flank the region to be copied. A 38-kb segment that carries genes ppsABCDE encoding five enzymes for antibiotic plipastatin synthesis and another genome locus as large as 100 kb including one essential gene were examined for positional cloning. The positional ...

background: tuberculosis (tb) is a major cause of death worldwide. finding an effective vaccine against tb is the best way to control it. several vaccines against this disease have been developed but none are completely protective. the aim of this study was to design and construct a cloning vector containing the mycobacterium tuberculosis (m. tuberculosis) heat shock protein x (hspx). methods: ...