Background and Aims: The aim of this study was cloning and expression of rabies virus glycoprotein by a eukaryotic expression plasmid pcDNA3.1(+) in BSR cell line. This construct might be used for a potential DNA vaccine. Materials and Methods: Glycoprotein gene was synthesized and cloned into pBluescript vector and then sub cloned into eukaryotic expression vector (pcDNA3.1(+)). After verifica...

We propose a novel method for direct cloning of foreign genes into baculoviruses which avoids the use of bacterial transfer vectors. The foreign gene to be inserted is derived by PCR using appropriate primers each of which contains an additional 50 nt of baculovirus sequence for homologous recombination between the PCR-derived DNA and the baculovirus DNA, thus accomplishing insertion of the for...

products. BioTechniques 12:28-30. 10.Scharf, S.J., G.T. Horn and H.A. Erlich. 1986. Direct cloning and sequence analysis of enzymatically amplified genomic sequences. Science 233:1076-1078. 11.Shuldiner, A.R., L.A. Scott and J. Roth. 1990. PCR-induced (ligase free) subcloning: a rapid reliable method to subclone polymerase chain reaction (PCR) product. Nucleic Acids Res. 18:1920. 12.Starr, L. a...

PREMISE OF THE STUDY Positional cloning in combination with phenotyping is a general approach to identify disease-resistance gene candidates in plants; however, it requires several time-consuming steps including population or fine mapping. Therefore, in the present study, we suggest a new combined strategy to improve the identification of disease-resistance gene candidates. METHODS AND RESULT...

BACKGROUND The advent of genomics-based technologies has revolutionized many fields of biological enquiry. However, chromosome walking or flanking sequence cloning is still a necessary and important procedure to determining gene structure. Such methods are used to identify T-DNA insertion sites and so are especially relevant for organisms where large T-DNA insertion libraries have been created,...

BACKGROUND The combined use of restriction enzymes with PCR has revolutionized molecular cloning, but is inherently restricted by the content of the manipulated DNA sequences. Uracil-excision based cloning is ligase and sequence independent and allows seamless fusion of multiple DNA sequences in simple one-tube reactions, with higher accuracy than overlapping PCR. RESULTS Here, the addition o...

Background Cystic fibrosis (CF), caused by mutations in the CF transmembrane conductance regulator gene, is a common autosomal recessive disease. Accurate isolation and identification of the bacteria underlying these infections are is critical to the therapeutic management of CF. Objective To compare phenotypic bacterial identification with a molecular method in a CF patient sputum. Methods...

AIM Idiotype, the unique part of immunoglobulin molecule expressed on the surface of B-cells, represents a specific antigen for vaccination against lymphoma. We have developed a rapid method for immunoglobulin variable fragments cloning, assembling and expression of recombinant idiotype protein in Escherichia coli. METHODS PCR with specially designed panel of primers was used for direct ampli...

colonization of nontypeable haemophilus influenzae (nthi) in nasopharynx causes respiratory tract disease. in 80% of clinical isolates, hmw proteins are the major adhesions and induce protective antibodies in the hosts. therefore, it can be used as a vaccine candidate. the aim of this study is designing and cloning of the conserved regions of nthi hmw1 core binding domain.in this study, the sta...