ABSTRACT
We hereby present the complete sequence and annotation of pRG930cm, a spectinomycin/streptomycin/chloramphenicol-resistant cosmid vector. pRG930cm (17,256 bp; GenBank Accession No.: FM174471) has a broad host range, and is stably maintained by a number of Gram-negative bacteria including Pseudomonas spp, Escherichia coli, Agrobacterium tumefaciens and Azorhizobium caulinodans ORS571. pRG930cm is already widely used and its sequence will aid efficient construction and analysis of cosmid libraries.
Subject(s)
Azorhizobium caulinodans/genetics , Cosmids , Escherichia coli/genetics , Pseudomonas/genetics , Agrobacterium tumefaciens/genetics , Chloramphenicol Resistance , Genetic Engineering , Spectinomycin , StreptomycinABSTRACT
Objective:To construct a recombinant adenovirus containing cytosine deaminase( CD ) gene and thymidine kinase( TK ) fusion gene for the gene therapy research of malignant tumors. Methods: A recombinant cosmid containing CD and TK fusion gene was constructed, and then mixed with DNA TPC and co transfected to the 293 cells by calcium phosphate coprecipitation. Results: The results of restriction and PCR showed the insertion was right and the recombinant adenovirus generated contained the CD and TK fusion gene without replication competent adenovirus. Conclusion: The recombinant adenovirus generated is E1 and E3 deleted and contains double suicide gene needed, which can be further studied for gene therapy.