BACKGROUND Cloning vectors capable of retroviral transduction have enabled stable gene overexpression in numerous mitotic cell lines. However, the relatively small number of feasible restriction enzyme sequences in their cloning sites can hinder successful generation of overexpression constructs if these sequences are also present in the target cDNA insert. RESULTS Utilizing ligation-independ...

Vector construction with restriction enzymes (REs) typically involves the ligation of a digested donor fragment (insert) to a reciprocally digested recipient fragment (vector backbone). Creating a suitable cloning plan becomes increasingly difficult for complex strategies requiring repeated insertions such as constructing multiple short hairpin RNA (shRNA) expression vectors for RNA interferenc...

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...

Simple and low-cost recombinant enzyme-free seamless DNA cloning methods have recently become available. In vivo Escherichia coli cloning (iVEC) can directly transform a mixture of insert and vector DNA fragments into E. coli, which are ligated by endogenous homologous recombination activity in the cells. Seamless ligation cloning extract (SLiCE) cloning uses the endogenous recombination activi...

background: laron syndrome, is a disease that treated by igf-1. this protein is a single chain and has three disulfide bonds. people with laron syndrome have low rates of cancer and diabetes, although they appear to be at increased risk of casual death due to their stature. igf-1 is synthesized by many tissues and is secreted from liver as an endocrine hormone which is transmitted to other tiss...

Yeast-based in vivo cloning is useful for cloning DNA fragments into plasmid vectors and is based on the ability of yeast to recombine the DNA fragments by homologous recombination. Although this method is efficient, it produces some by-products. We have developed an "ultra-low background DNA cloning system" on the basis of yeast-based in vivo cloning, by almost completely eliminating the gener...

Background & objective:  Tuberculosis (TB) remains a major cause of death around the world. Bacillus Calmette Guérin (BCG) is the only vaccine used in TB prevention that has a protective effect in children, but its effectiveness declines in adults. Design and development of new vaccines is the most effective way against TB. The aim of this study was to design and construc...