Plasmid cloning vectors

Schmidhauser, T. J., Ditta, G. Helinski, D. R. (1988). Btoad-host-range plasmid cloning vectors for gram-negative bacteria. In Vectors A Survey of Molecular Cloning Vectors and their Uses, ed. R. L. Rodriguez D. T. Denhardt, pp. 287-332. Boston Butterworth. 

Prepare a plasmid cloning vector, preferably choosing one that contains the same restriction site as the one used for the genomic DNA digest so that the insert may be excised cleanly from the plasmid for subsequent... 

Cloning of eukaryotic DNA into a plasmid cloning vector. 

Plasmid cloning vector cleaved with EcoSl and PvuB. 

Mather, M.W. and Fee, J.A. (1992) Development of plasmid cloning vectors for Ihermus thermophilus HB8 expression of a heterologous, plasmid-borne kanamycin nucleotidyltransferase gene. Appl. Ertvirort. Microbiol, 58, 421-425. 

Bierman M, Logan R, O Brien K, Seno ET, Rao RN, Schoner BE. Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia colt to Streptomyces spp. Gene 1992 116 43-39. 

Kuhstoss S, Richardson MA, Rao RN. Plasmid cloning vectors chat integrate site-specibcally in Streptomjees spp. Gene 1991 97 143-146. 

Bacterial plasmids are small, circular, duplex DNA molecules whose natural function is to confer antibiotic resistance to the host cell. Plasmids have several properties that make them extremely useful as cloning vectors. They exist as single or multiple copies within the bacterium and replicate independently from the bacterial DNA. The complete DNA sequence of many plasmids is known hence, the precise location of restriction enzyme... 

Fig. 24.1 The requirement for a cloning vector (A) fragments of DNA introduced into the hacterium hy transformation do not undergo replication and gradually are diluted out of the population (B) DNA fragments introduced into plasmids are inherited hy both daughter progeny cell division.

Plasmid An extracbromosomal circular DNA molecule in bacteria. Often used as cloning vector. 

Figure 4.4. Schematic illustration of directional topoisomerase cloning of PCR products into the pUNI vector. The PCR product to be cloned has the sequence 5 -CACC appended at the 5 end to direct the orientation of cloning. The Vaccinia virus topoisomerase I enzyme forms a covalent adduct with the cloning vector to create a cloning competent plasmid construct. The loxP site is 5 to the insertion site. The vector and PCR product are designed to fuse the ORF in-frame with loxP.

Integration of the various fragments generated into cloning vectors, which are themselves small DNA molecules capable of self-replication. Typically, these are plasmids or viral DNAs and the composite or engineered DNA molecules generated are called rDNA. 

An essential feature of the cloning vector used is that it must be capable of self-replication in the cell into which it is introduced, which is usually E. coli. Two of the most commonly used types of vector in conjunction with E. coli are plasmids and bacteriophage X. Plasmids are circular extra-chromosomal DNA molecules, generally between 5000 and 350 0000 bp in length, that are found naturally in a wide range of bacteria. They generally house several... 

---