Plasmid vectors are convenient for cloning of small dna fragments for restriction mapping andfor studying regula. A cosmid is a type of hybrid plasmid that contains a lambda phage cos sequence. A cloning vector is a small piece of dna, taken from a virus, a plasmid, or the cell of a higher organism, that can be stably maintained in an organism, and into which a foreign dna fragment. A cloning vector is a small piece of dna that can be stably maintained in an organism, and into which a foreign dna fragment can be inserted for cloning. This article throws light upon the three types of cloning vectors used in recombinant dna technology. Bacteriophage virus infecting bacteria are called bateriophage. Extensive research has been directed toward the development of multipurpose lambda vectors for cloning ever since the potential of using coliphage lambda as a cloning vector.
Lambda as a cloning vector current protocols wiley. Schematic representation of the pbr322 plasmid, one of the first plasmids widely used as a cloning vector. A cloning vector is a small piece of dna that can be stably maintained in an organism, and into which a foreign dna fragment can be inserted for cloning purposes. Bacteriophage lambda cloning vectors sciencedirect. Mke phage is a suspension of infectious virions derived from the ph. Although cloning in cosmids is preferred for certain spe. Bacteriophage lambda vectors we have talked about plasmids as vectors for cloning small pieces of dna. The transfer of genetic elements from one bacterium to another by a bacteriophage is termed as transduction. A history of molecular cloning 1971 restriction enzyme.
Cloning vector based on lambda bacteriophage part2. Viruses and bacteriophages the viruses that infect bacteria provided vectors for some of the first cloning experiments, and they continue to be used in a variety of applications. Therefore size of the gene of interest is limited in plasmid cloning vectors. Bacteriophage lambdabased expression vectors springerlink. Lambda fix is similar to lambda dash, except that it allows cloning into a partially filledin xhol site as described in the text.
Cloning and sequencing of hfq host factor required for synthesis of bacteriophage q beta rna gene of salmonella typhimurium isolated from poultry parthasarathi behera, muhammed kutty, bhaskar sharma, ajay kumar and meeta saxena. Cloning vectors cloning vectors are dna molecules that are used to transport cloned sequences between biological hosts and the test tube. Genetic organization of lambda dna the lambda phage is a typical example of bacteriophage which has a polyhedral head. Cosmid vectors are developed by combining the features of the plasmid vector and the bacteriophage vector. Therefore, lambda phages are mostly used as a cloning vector.
V m chauthaiwale, a therwath, and v v deshpande division of biochemical sciences, national chemical laboratory, pune, india. Origin of replication, multiple cloning site and selectable marker are obtained from the plasmid and only the cohesive site or cos site region is taken from lambda phage. Genetic organization of lambda dna the lambda phage is a typical example of bacteriophage which has a polyhedral head and tail. M phage has been variously modified to give rise to a mp mp series of cloning vectors which can be used for cloning.
The general steps of gene cloning with any vector are as follows figure 1. A vector is used to amplify a single molecule of dna into many copes. Their construction incorporates mutations that make them simple to use and also greatly increases their safety for the biological containment of cloned recombinant dna. A cloning vector or cloning vehicle is a dna molecule in which foreign dna can be inserted or integrated. A history of molecular cloning 1971 restriction enzyme 1977 report ot the first 1975 cloning vector launch rebase lg91 introduction user cloning 36 2007 introduction ot ligation independent cloning. Virus that infect bacteria is known as bacteriophage.
Three of the charon vector phages, 3a, 4a, and 16a, have been certified for use as ek2 vector. The charon lambda bacteriophages have been developed as vectors for cloning. The insertion of dna fragment in to the cloning vector is carried out by treating the vehicle and the foreign dna with a restriction enzyme that create the same overhang. D herelle coined the term bacteriophage meaning bacterial eater to describe the agents bacteriocidal activity. Principles of cloning, vectors and cloning strategies. This virus is temperate and may reside within the genome of its host through lysogeny.
A simple method for generating phage collections representing eukaryotic genomes has been developed by using a novel bacteriophage lambda vector, lambda 1059. The efficiency of packaging and infection, and the simplicity of plaque screening are advantages of lambda as a cloning vector. But the plasmid cloning vectors which carry more than 5 to 10 kb gene of interest are often unstable. This molecule is further capable of replicating within host cell to produce multiple clones of recombinant dna. When a single vector containing a single dna fragment is introduced into a host cell, large numbers of this fragment are reproduced along with the vector. We have developed a bacteriophage a cloning vector, xorf8, that can be used for the construction of cdna libraries. Bacteriophage lambda has been in use as a cloning vector for over 25 years, and has been used extensively as an expression vector.
Cloning and sequencing of hfq host factor required for. Cloning vector is a dna molecule that carries foreign dna into a host cell, replicates inside a. The vector phage may be a useful control for phage elisa or for titering of phage stocks. The bacteriophage known as m forms the basis of cloning systems designed to easily introduce mutations into genes inserted into the phage genome. The first page of the pdf of this article appears above.
Lytic pathway large scale production of bacteriophage particles that eventually leads to the lysis of the cell. For cloning, the isolated replicative form of mke is available as part of the ph. The limitation of this vector is the size of dna that can be introduced into the cell by. Cloning vectors used in recombinant dna technology. Cloning vectors the two molecules that are required for cloning are the dna to be cloned and a cloning vector. We have talked about plasmids as vectors for cloning small pieces of dna. Bacteriophage a cloning system for the construction of.
This chapter will trace the development of a versatile series of vectors for different cloning problems. The limitation of this vector is the size of dna that. A lambda insertion type cdna cloning vector, lambda zap, has been constructed. These are fused together to obtain the cosmid vector. Twort in great britian 1915 and felix d herelle in france1917. Bacteriophage and viral cloning vectors springerlink. The most important bacteriophages have been the filamentous phages especially m, bacteriophage lambda, and bacteriophage. Bacteriophage lambda as a cloning vector microbiology. M phage with lacz containing multiple cloning site.
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