A new versatile mammalian vector system for protein production, cell biology analyses, and cell factory engineering was developed. The vector system applies the ligation-free uracil-excision based technique--USER cloning--to rapidly construct mammalian expression vectors of multiple DNA fragments and with maximum flexibility, both for choice of vector backbone and cargo. The vector system inclu...

Background: Human epidermal growth factor receptor 2 (HER2) is over-expressed in breast, ovarian, gastric, and prostate cancers used as a tumor marker the diagnosis of cancer. Monoclonal antibodies have been diagnostic therapeutic tool against HER2. Because difficulties associated with stability complexity construct high cost antibody production, we aimed to investigate, cloning, expression HER...

Vast numbers of proteins work cooperatively to exert their functions in various cells. In order to understand the functions and molecular mechanisms of these proteins in plants, analyses of transgenic plants that concomitantly express two protein-coding genes are often required. We developed a novel Gateway cloning technology-compatible binary vector system, the R4 dual-site (R4DS) Gateway clon...

Many different strategies are used for cloning polymerase chain reaction (PCR) products. Some use restriction sites pre-integrated into primers or contained in a generated fragment. Others, such as the one used by Stratagene (La Jolla, CA, USA) in its pCR-Script Direct SK(+) Cloning Kit, are based on a blunt-end ligation. These strategies require the use of additional enzymes to polish the end...

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...

background and objective: the outer membrane protein w ( ompw ) of vibrio cholerae is involved in stimulating the im- mune response via induction of protective immunity. it also plays an important role in bacterial pathogenesis by increasing the adaptability of pathogenic strains. in this study we aimed to clone v. cholerae ompw gene in the strain x-33 of pichia pastoris. materials and methods:...

We have constructed a small and highly efficient binary Ti vector pSiM24 for plant transformation with maximum efficacy. In the pSiM24 vector, the size of the backbone of the early binary vector pKYLXM24 (GenBank Accession No. HM036220; a derivative of pKYLX71) was reduced from 12.8 kb to 7.1 kb. The binary vector pSiM24 is composed of the following genetic elements: left and right T-DNA border...

The 22-KDa is the protein product of human growth hormone (hGH) gene isoform 1 of the pituitary hormone (Ahmed et. al., 1993). hGH gene was amplified with PCR from human growth hormone cDNA clone by specific designed primers containing extraterminals having Nco I and Bam HI restriction sites. The PCR product obtained was analyzed by enzyme digestion and DNA sequencing. The product was cloned in...

The bacteriophage P1 cloning system can package and propagate DNA inserts that are up to 95 kilobases. Clones are maintained in Escherichia coli by a low-copy replicon in the P1 cloning vector and can be amplified by inducing a second replicon in the vector with isopropyl beta-D-thiogalactopyranoside. To overcome the necessity of screening clones for DNA inserts, we have developed a P1 vector w...