BACKGROUND In high-throughput demanding fields, such as biotechnology and structural biology, molecular cloning is an essential tool in obtaining high yields of recombinant protein. Here, we address recently developed restriction-free methods in cloning, and present a more cost-efficient protocol that has been optimized to improve both cloning and clone screening. RESULTS In our case study, t...

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...

chicken anemia virus was detected by pcr in tissue samples collected from poultry flocks in gujarat,india. the vp1, vp2 and vp3 gene sequences of cav from anand, gujarat were obtained after cloning thepcr products in pdrive cloning vector. nucleotide sequence alignment with other cav sequencesdemonstrated overall identity of 95-98.8%, 98.8-99.8% and 98.8-100% for vp1, vp2 and vp3 regions,respec...

Direct cloning of PCR fragments by TA cloning or blunt end ligation are two simple methods which would greatly benefit high-throughput (HTP) cloning constructions if the efficiency can be improved. In this study, we have developed a ribosomal binding site (RBS) switching strategy for direct cloning of PCR fragments. RBS is an A/G rich region upstream of the translational start codon and is esse...

Rice (Oryza sativa) is a staple food consumed by the majority of world’s population. Climate change, however, has created significant threat to our security as it posed severe effects on rice production. The emergence genome editing technology opened new era in crop improvement. Hence, this study aims develop CRISPR/Cas9 construct drought tolerance for O. sativa subsp. indica cv. IR64 using Gol...