Phage cloning vectors

DA Hopwood, MJ Bibb, KF Chater, T Kieser. Plasmid and phage vectors for gene cloning and analysis in Streptomyces. Meth Enzymol 153 116-167, 1987. 

Cloning DNA sequences into plasmid and phage vectors... 

Rapid random sequencing by shotgun cloning into single stranded phage vectors (Sanger et al., 1980)... 

Anderson, S., M.J. Gait, L. Mayol and I.G. Young, 1980. A short primer for sequencing DNA cloned in the single-stranded phage vector M13mp2. Nucleic Acids Res. 8, 1731. 

In our first attempts to clone the most important allergens of the two molds, cDNA libraries were prepared in phage vectors and standard immunological... 

Fig. 1, Organization of cab genes in nuclear DNA from petunia and tomato, showing the tandem and head-to-head arrangement of the genes. Each region of DNA represents a restriction fragment of nuclear DNA cloned in a A phage vector. Redrawn from Dunsmuir et al, [28] and Pichersky et al. [30],...

In some circumstances the introduction of a cloning vector into a host cell is a trivial process. For example, phage vectors are designed so they introduce recombinant DNA in an infective process called transfection, and some bacteria take up plasmids unaided. However, most host cells must be induced to take up foreign DNA. Several methods are used. In some prokaryotic and eukaryotic cells, the addition of Ca2+ to the medium promotes uptake. In others, a process called electroporation, in which cells are treated with an electric current, is used. One of the most effective methods for transforming animal and plant cells is the direct microinjection of genetic material. Transgenic animals, for example, are created by the microinjection of recombinant DNA into fertilized ova. 

In cDNA cloning, expressed mRNAs are reverse-transcribed into complementary DNAs, or cDNAs. By a series of reactions, single-stranded cDNAs are converted Into double-stranded DNAs, which can then be ligated into a phage vector (see Figure 9-15). 

Bacterial plasmids and X phage serve as cloning vectors. Describe the essential features of a plasmid and a X phage vector. What are the advantages and applications of plasmids and X phage as cloning vectors ... 

---