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    MSC Biotech 02sem ProfBhuminathTripathi

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    Atashi Mandal
    The document discusses various types of cloning vectors such as plasmids, bacteriophages, BACs, YACs, and HACs that are used to artificially carry foreign DNA into host cells. It describes the essential characteristics of cloning vectors including self-replication, unique restriction sites, and marker genes. The different vectors each have advantages and limitations depending on the size of DNA insert and intended application.

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    MSC Biotech 02sem ProfBhuminathTripathi

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    Atashi Mandal
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    The document discusses various types of cloning vectors such as plasmids, bacteriophages, BACs, YACs, and HACs that are used to artificially carry foreign DNA into host cells. It describes the essential characteristics of cloning vectors including self-replication, unique restriction sites, and marker genes. The different vectors each have advantages and limitations depending on the size of DNA insert and intended application.

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    The document discusses various types of cloning vectors such as plasmids, bacteriophages, BACs, YACs, and HACs that are used to artificially carry foreign DNA into host cells. It describes the essential characteristics of cloning vectors including self-replication, unique restriction sites, and marker genes. The different vectors each have advantages and limitations depending on the size of DNA insert and intended application.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    35 views6 pages

    MSC Biotech 02sem ProfBhuminathTripathi

    Uploaded by

    Atashi Mandal
    The document discusses various types of cloning vectors such as plasmids, bacteriophages, BACs, YACs, and HACs that are used to artificially carry foreign DNA into host cells. It describes the essential characteristics of cloning vectors including self-replication, unique restriction sites, and marker genes. The different vectors each have advantages and limitations depending on the size of DNA insert and intended application.

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    M.Sc.

    (Biotechnology) II Semester
    Paper: Genetic Engineering, Genomics and Proteomics
    By: Professor Bhumi Nath Tripathi
    (Whatsapp: 9694218853; Email: bhuminath.tripathi@igntu.ac.in)

    Stay at home, stay safe. Your life is precious for the nation.

    Cloning Vectors
    • Genetic vectors are vehicles for delivering foreign DNA into recipient cells.
    • In molecular cloning, a vector is a DNA molecule used as a vehicle to
    artificially carry foreign genetic material into another cell, where it can
    be replicated and/or expressed.
    • Vectors can replicate autonomously and typically include features to
    facilitate the manipulation of DNA as well as a genetic marker for their
    selective recognition.
    • The different types of vectors available for cloning are plasmids,
    bacteriophages, bacterial artificial chromosomes (BACs), yeast
    artificial chromosomes (YACs) and mammalian artificial
    chromosomes (MACs).
    • The cloning vectors are limited to the size of insert that they can carry.
    Depending on the size and the application of the insert the suitable vector
    is selected for a particular purpose.
    Essential Characteristics of Cloning Vectors
    Regardless of the selection of a vector, all vectors are carrier DNA molecules.
    These carrier molecules should have few common features in general such as:
    • It must be self-replicating inside host cell.
    • It must possess a unique restriction site for RE enzymes.
    • Introduction of donor DNA fragment must not interfere with replication
    property of the vector.
    • It must possess some marker gene such that it can be used for later
    identification of recombinant cell (usually an antibiotic resistance gene that
    is absent in the host cell).
    • They should be easily isolated from host cell.

    Plasmids
    • Plasmids are extra chromosomal circular double stranded DNA replicating
    elements present in bacterial cells.
    • Plasmids show the size ranging from 5.0 kb to 400 kb.
    • Plasmids are inserted into bacterial calls by a process called
    transformation.
    • Plasmids can accommodate an insert size of upto 10 kb DNA fragment.
    • Generally, plasmid vectors carry a marker gene which is mostly a gene for
    antibiotic resistance; thereby making any cell that contains the plasmid will
    grow in presence of the selectable corresponding antibiotic supplied in the
    media.

    Bacteriophage

    • The viruses that infect bacteria are called bacteriophage. These are
    intracellular obligate parasites that multiply inside bacterial cell by making
    use of some or all of the host enzymes.
    • Bacteriophages have a very high significant mechanism for delivering its
    genome into bacterial cell. Hence it can be used as a cloning vector to
    deliver larger DNA segments.
    • Most of the bacteriophage genome is non-essential and can be replaced
    with foreign DNA.
    • Using bacteriophage as a vector, a DNA fragment of size up to 20 kb can
    be transformed.

    Bacterial artificial chromosomes (BACs)


    • Bacterial artificial chromosomes (BACs) are simple plasmid which is
    designed to clone very large DNA fragments ranging in size from 75 to 300
    kb.
    • BACs basically have marker like sights such as antibiotic resistance genes
    and a very stable origin of replication (ori) that promotes the distribution of
    plasmid after bacterial cell division and maintaining the plasmid copy
    number to one or two per cell.
    • BACs are basically used in sequencing the genome of organisms in
    genome projects (example: BACs were used in human genome project).
    • Several hundred thousand base pair DNA fragments can be cloned using
    BACs.

    Yeast artificial chromosomes (YACs)


    • YACs are yeast expression vectors.
    • A very large DNA fragments whose sizes ranging from 100 kb to 3000 kb
    can be cloned using YACs.
    • Mostly YACs are used for cloning very large DNA fragments and for the
    physical mapping of complex genomes.
    • YACs have an advantage over BACs in expressing eukaryotic proteins that
    require post translational modifications.
    • But, YACs are known to produce chimeric effects which make them less
    stable compared to BACs.

    Human artificial chromosomes (HACs)


    • Human artificial chromosomes (HACs) or mammalian artificial
    chromosomes (MACs) are still under development.
    • HACs are micro-chromosomes that can act as a new chromosome in a
    population of human cells.
    • HACs range in size from 6 to 10 Mb that carry new genes introduced by
    human researchers.
    • HACs can be used as vectors in transfer of new genes, studying their
    expression and mammalian chromosomal function can also be elucidated
    using these micro-chromosomes in mammalian system.
    Other Types of Vectors
    All vectors may be used for cloning and are therefore cloning vectors, but there
    are also vectors designed specially for cloning, while others may be designed
    specifically for other purposes, such as transcription and protein expression.

    1. Expression Vectors
    Vectors designed specifically for the expression of the transgene in the target cell
    are called expression vectors, and generally have a promoter sequence that
    drives expression of the transgene. Expression vectors produce proteins through
    the transcription of the vector’s insert followed by translation of
    the mRNA produced.

    2. Transcription Vectors
    Simpler vectors called transcription vectors are only capable of being transcribed
    but not translated: they can be replicated in a target cell but not expressed, unlike
    expression vectors. Transcription vectors are used to amplify their insert.

    Uses of Vectors
    Vectors have been developed and adapted for a wide range of uses. Two
    primary uses are:
    (1) to isolate, identify and archive fragments of a larger genome
    (2) to selectively express proteins encoded by specific genes.
    Vectors were the first DNA tools used in genetic engineering, and continue to be
    cornerstones of the technology.

    References
    1. https://nptel.ac.in/courses/102103017/pdf/lecture%2035.pdf
    2. https://www.nature.com/subjects/genetic-vectors
    3. https://www.sciencedirect.com/topics/agricultural-and-biological-sciences/vector-
    molecular-biology
    4. https://www.britannica.com/science/recombinant-DNA-technology#ref964476
    5. https://en.wikipedia.org/wiki/Vector_(molecular_biology)

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