Cloning Vectors for Recombinant DNA, Random DNA, cDNA Segments, Organo Chemically, Bacteriophages, Cosmid Phagemix

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Cloning and Expression Vectors Cloning and Expression Vectors Introduction Cloning Vectors for Recombinant DNA Plasmids as Vectors Three Phases of the Development of Plasmid Cloning Vectors Alpha-Complementation for Screening Using "blue-white"Clonies Nonsense Suppressors to Complement Termination Codons in Vectors E.Coli Plasmid Vectors pBR322 and pBr327 Vectors pUC Vectors pSP64-pSP65 pGEM-3, pGEM-4, pGEM-3Z, pGEM-4Z ΠAN13 from ΠAN7 Yeast Plasmid Vectors Retriever Vectors Agrobacterium Plasmid Vectors Lambda Phage (λ)Vectors Lambda Phage (Lysis and Lysogeny) Genetic Markers for the Selection and Screening of λ Vectors Development of Lambda (λ)Phage Vectors Insertion λ Vectors Replacement λ Vectors M13 Phage as a Vector for DNA Sequencing M13-vectors-of-mp-Series Hosts for M13 Vectors Cosmids as Vectors Phagemids as Vectors Phagemids pUC118 and pUC119 Phagemid pBluesript IIKS

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Cloning Vectors for Recombinant DNA

One of the most important uses of recombinant DNA technology is the cloning of (i) random DNA or cDNA segments, often used as probes or (ii) specific genes, which may be either isolated from the genome or synthesized organo-chemically or in the form of cDNA from mRNA.

This cloning of DNA is possible only with the help of another DNA molecule, which is capable of replicating in a host. This other DNA molecule is often used in the form of a vector, which could be a plasmid, a bacteriophage, a derived cosmid or phagemid (see later in this section), a transposon, a virus or even an artificial chromosome.

Techniques should also be available, which will allow selection of chimeric genomes obtained after insertion of foreign DNA from a mixture of chimeric and the original vector (see later). Another critical desired feature of any cloning vector is that it should possess a site at which foreign DNA can be inserted without disrupting any essential function.

Therefore, in each case an enzyme will also have to be selected which will cause a single break. Sometimes vectors are modified by inserting a DNA segment to create unique cleavage site(s) for one or more enzymes to facilitate its use in gene cloning. This inserted DNA with restriction sites for several enzymes is sometimes called a polylinker.

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BACs and PACs as Vectors for Cloning large DNA Segments (100-300 kb) Bacterial Artificial Chromosomes (BACs) P1-derived Artificial Chromosomes (PACs) Plant and Animal viruses as Vectors Plant Viruses (Cauliflower Mosaic virus or CaMV and Geminiviruses) Animal Viruses Transposons as Vectors Transposons of Higher Plants (Ac/Dc or Mul of Maize) Transposons of drosophila (P elements) YAC and MAC Vectors for Cloning very large DNA Segments (1000 kb) Yeast Artificial Chromosomes (YACs) Mammalian Artificial chromosomes (MACs) Expression Vectors Promoters Nopaline Synthase (nos) Promoter from T-DNA Dual Promoter of Mannopine Synthase (mas) genes 1 and 2 35S Promoter of CaMV Polyhedrin Promoter from Baculovirus Expression Cassettes Baculovirus and Expression Vector System for Insect Cells Virus Expression Vectors for Mammalian Cells Binary and Shuttle Vectors