Plasmid pSUP5011 contains a pMBl origin of replication, an origin of transfer (oriT), and genes encoding resistance to the antibiotics ampicillin, chloramphenicol and kanamycin. pSUP5011 was conjugally mobilized from Escherichia coli into the non-heterocystous, filamentous, nitrogen-fixing cyanobacterium Plectonema boryanum UTEX 594 in the presence of a helper plasmid, RP4. Transconjugant cyano...

the genome of hb v virus of serotype ayw cloned in pbr322 and expression shuttle vector p yes2 were used for construction of the hbsag chimeric genes and their expression in saccharomyces cerevisiae. two recombinant plasmids were constructed. one of them contained the coding sequences for the major polypeptide of surface antigen. another construct carried the major polypeptide with the pre-s2 a...

Actinobacillus actinomycetemcomitans, a periodontal pathogen, has been strongly implicated in human periodontal disease. Advances in the molecular analysis of A. actinomycetemcomitans virulence factors have been limited due to the unavailability of systems for genetic transfer, transposon mutagenesis, and gene complementation. Slow progress can be traced almost exclusively to the lack of gene v...

Phospholipase Cg2 (PLCg2) induces apoptosis of immune and tumor cells; however, it remains unclear whether PLCg2 promotes hepatocyte apoptosis during liver regeneration (LR). Therefore, to establish a framework for further exploring the function of PLCg2, we generated recombinant adenoviruses carrying a template encoding short hairpin (sh)-RNA targeting PLCg2 (Ad-PLCg2-shRNA), which were used t...

odv-e25(e25) is one of the core genes of baculoviruses. To investigate how it functions in the replication cycle of a baculovirus, a number of Autographa californica multiple nucleopolyhedrovirus recombinants with e25 under control of the promoter of immediate early gene ie1, or the promoter of the very late hyperexpressed gene p10, were constructed using a bacmid system, and the effects of ear...

Bacterial artificial chromosome (BAC) mediated transgenesis has proven to be a highly reliable way to obtain accurate transgene expression for in vivo studies of gene expression and function. A rate-limiting step in use of this technology to characterize large numbers of genes has been the process with which BACs can be modified by homologous recombination in Escherichia coli. We report here a ...

BioTechniques 625 Adenoviral shuttle vectors to express short hairpin RNAs (shRNA) using RNA polymerase III (RNA pol III) promoters (mouse U6, human H1, or human U6) have been constructed in several laboratories including Welgen (Worcester, MA, USA), BD Biosciences (San Jose, CA, USA), Invitrogen (Carlsbad, CA, USA), and GeneScript (Piscataway, NJ, USA). Welgen and Invitrogen developed adenovir...

Here we describe a convenient method to generate homologous recombinant baculoviral genomes in E. coli. The recombination takes place with the aid of recombination enzymes provided by the phage lambda Red system between a bacmid (a baculoviral genome that can replicate in bacteria) and a linear fragment. Proof of concept was provided when the cathepsin gene (v-cath) of the Helicoverpa armigera ...