The gene encoding MPB51 was amplified from M. bovis Valleel11 chromosomal DNA using PCR technique, and the PCR product was approximately 800 bp DNA segment. Using T-A cloning technique, the PCR product was cloned into pGEM-T vector and cloning plasmid pGEM-T-51 was thus constructed successfully. pGEM-T-51 and pET28a( + ) were digested by BamH I and EcoR I double enzymes. The purified MPB51 gene...

The surface layer protein encoding genes from five mosquito-pathogenic Bacillus sphaericus isolates were amplified and sequenced. Negative staining of the S-layer protein extracted from the cell wall of wild-type B. sphaericus C3-41 was prepared. It showed a flat-sheet crystal lattice structure. Two genes encoding the entire and N-terminally truncated S-layer protein (slpC and DeltaslpC respect...

We amplified S14R protein gene cDNA of porcine, cloned it into a prokaryotic expression plasmid, and expressed it in Escherichia coli. A pair of primers was designed based on the cDNA sequence of the porcine S14R gene in GenBank. The target gene fragment from porcine liver tissue was amplified by RT-PCR. Confirmed by auto-sequencing, the target gene fragment was subcloned into an expression vec...

BACKGROUND AND OBJECTIVES In present study we aimed to clone the luxI gene encoding N-acyl-homoserine synthase detected in clinical isolates of Acinetobacter baumannii and study its expression in Escherichia coli transformants. MATERIALS AND METHODS Four A. baumannii hospital strains which demonstrated strong biofilm activity were selected in this investigation. The presence of luxI gene was ...

AIM To study immunogenicity of Pseudomonas N terminal flagellin as an adjuvant for Acinetobacter baumannii (A. baumannii) biofilm associated protein (Bap). METHODS The N terminal flagellin gene was amplified. The pET28a (+) and polymerase chain reaction products were digested with HindIII and EcoR I. The ligation of N terminal flagellin into pET28a (‏+) was performed using T4 DNA ligase and w...

Metallothioneins (MTs) are low molecular weight cysteine-rich proteins that bind to metals. Owing their high cysteine (Cys) content, MTs effective mediators of heavy metal detoxification. To enhance the binding ability MT from freshwater crab Sinopotamon henanense (ShMT), sequence-based multiple sequence alignment (MSA) and structure-based docking simulation (MDS) were conducted in order identi...

To obtain the recombinant group 2 allergen product of Dermatophagoides farinae (Der f 2), the Der f 2 gene was synthesized by RT-PCR. The full-length cDNA comprised 441 nucleotides and was 99.3% identical to the reference sequence (GenBank AB195580). The cDNA was bound to vector pET28a to construct plasmid pET28a(+)-Der f 2, which was transformed into E. coli BL21 and induced by IPTG. SDS-PAGE ...

Antibody Production The cDNA sequence corresponding to amino acids 1-416 of Vha100-1 was amplified by PCR to incorporate BamHI and NotI restriction sites and then cloned into to the pET28a (Invitrogen) vector for protein expression. The primers used were: 5’-CGGAATTCATGGGTTCCCTATTCCGCAGCGAGG-3’ and 5’-CCGCTCGAGGGATTCATCTCCCGATAGCTGGCC-3’. Guinea pig antibodies against this domain were raised by...