Bacteriophage construction

Fig. 4. Construction of recombinant phage in vectors derived from bacteriophage lambda where E represents the enzyme EcoRl. Other terms are defined...

Expression vectors are engineered so that any cloned insert can be transcribed into RNA, and, in many instances, even translated into protein. cDNA expression libraries can be constructed in specially designed vectors derived from either plasmids or bacteriophage A. Proteins encoded by the various cDNA clones within such expression libraries can be synthesized in the host cells, and if suitable assays are available to identify a particular protein, its corresponding cDNA clone can be identified and isolated. Expression vectors designed for RNA expression or protein expression, or both, are available. 

Yeast (Saeeharomyees eerevisiae) has a genome size of 1.21 X 10 bp. If a genomic library of yeast DNA was constructed in a bacteriophage A vector capable of carrying 16-kbp inserts, how many indi-... 

FIGURE 6.3 Construction of a brain-specific cDNA library. Messages (mRNA) are isolated from a tissue of interest. In this case, the target tissue is the brain. The mRNAs are converted to cDNA and each cDNA is inserted into the DNA of the bacteriophage lambda. The end result is a brain-specific cDNA library. 

XEMBL3A Bacteriophage E. coli Genomic library construction. Reduced chance of escape from laboratory biological containment... 

The transfer of resistance can be achieved by conjugation, transduction, and transformation. There is also a phenomenon of transposition by which resistance determinants pass from one plasmid to another or to a chromosome or to a bacteriophage, thus allowing construction of new plasmids under the pressure of new antibiotic exposure. 

Bacteriophage A vectors that accommodate foreign DNA fragments generated by a variety of restriction endonucleases have been constructed. The recombinant DNA molecules that incorporate some of these vectors can be introduced directly into E. coli by transfection. Alternatively, recombinant DNA molecules can be packaged into phage particles and subsequently infected into suitable host cells. 

Fig. 6.18. A flow chart for constructing genetic libraries using the bacteriophage expression vector Agtll (After Simpson et al., 1986.)...

Abrol S, Sampath A, Arora K, Chaudhary VK, Construction and characterization of Ml3 bacteriophages displaying gpl20 binding domains of human CD4, Indian J. Biochem. Biophys., 31 302-309, 1994. 

Christian RB, Zuckermann RN, Kerr JM, Wang L, Malcom BA, Simplified methods for construction, assessment and rapid screening of peptide libraries in bacteriophage, J. Mol. Biol., 227 711-718, 1992. 

Markland W, Roberts BL, Saxena MJ, Guterman SK, Ladner RC, Design, construction and function of a multicopy display vector using fusion to the major coat protein of bacteriophage M13, Gene, 109 13-19, 1991. 

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