DNA cloning in microbial plasmids is a convenient and versatile method of DNA amplification. Suitable plasmid vectors and efficient transformation systems, which are presently available for many micro-organisms, allow introduction and establishment ofrecombinant plasmidsgenerated via enzymatic reactionsin vitro...
Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of ...
Vector construction employs ligation and restriction techniques known in the art. The quantity of DNA available can be increased by cloning the desired fragments, ie, inserting into a suitable cloning vehicle, such as pBR322, pUC13 or pUC8, transforming and replicating in E. coli, and, optional...
The term “DNA sequence encoding a Factor VIII protein” as used herein means DNA which encodes a Factor VIII protein, i.e., such DNA may be a full-length gene encoding a full-length Factor VIII protein, or a truncated gene, or a mutated gene encoding a biologically active Factor VIII ...
2.3. Cloning and expression studies Taq DNA polymerase clone was generated by PCR using primers as indicated in Table 1. PCR amplification was carried out using pTrc99A-Taq polymerase plasmid as a template and the amplified product was ligated to Nde I/Hind III digested pET26b vector (Novagen...
Hartley JL, Temple GF, Brasch MA: DNA cloning using in vitro site-specific recombination. Genome Res. 2000, 10: 1788-95. 10.1101/gr.143000. Article CAS Google Scholar Hendrickson WA, Horton JR, LeMaster DM: Selenomethionyl proteins produced for analysis by multiwavelength anomalous diffraction ...
The results from the SoPIP1;2 selection emphasise the importance of evaluating the outcome of the antibiotic selection not only at the protein level, but also at the DNA level, to ensure that the selection is efficient and that satisfactory gene dosage is obtained. If, as in the case with...
Cloning of foreign genes like GCSF, PTH, SAK and MAP into p24MGFPm vector All the genes were PCR amplified using synthetic genes (procured from GenScript, USA and BioServe Technologies, India) as template with Taq DNA polymerase and the PCR products were cloned into p24MGFPm vector as NdeI...
In certain embodiments, the recombinant DNA constructs encoding the fusion proteins will contain a cloning site in place of DNA encoding one or more of the heterologous domains, thus permitting the practitioner to introduce DNA encoding a heterologous domain of choice. In some embodiments the kit ...
A temperature-inducible, ampicillin-resistant expression vector, pXAFB1, which is diagramed in FIG. 1, containing the DNA coding region for a protein of 388 amino acids, comprising a portion of fibronectin binding protein B from S. aureus, was transfected into E. coli strain MZ-1. The ...