This document discusses different types of cloning vectors, including plasmids, cosmids, and bacteriophages. Plasmids like pBR322 and pUC19 are commonly used plasmid vectors that are small in size, contain origins of replication and antibiotic resistance genes, and have restriction enzyme sites. Cosmids are hybrid vectors derived from plasmids that contain phage packaging sites. Bacteriophages like M13 and lambda are also used as vectors, as their viral DNA can transform bacterial cells and replicate inside them.
1. Cloning Vectors
For B.Sc III(Semester VI)
(As per the syllabus of S.G.B.A.U.
Amravati)
By : Mohanish V. Wankhade
M.Sc.(Botany),CSIR-NET,SET,GATE
2. Cloning Vectors
• Vectors are the carrier of gene from donor to the
host.
Salient features of good vector
• Vector should be small in size i.e. 10 kbp.
•It must possess origin of replication.
•It should possess several restriction sites.
•It should possess marker genes and antibiotic
resistant genes.
•It should possess a site at which foreign gene can be
inserted without disrupting any essential function.
3. Types of vectors
Plasmid as a vector
•Plasmid are the small circular, extra chromosomal
DNA found in bacteria.
•There are several plasmid vectors such as PBR
322,PUC ,Ti Plasmid, Ri Plasmid the plasmid vector
designated as P
i) pBR322 plasmid vector
• It is the first artificial cloning vector developed in
1977 by Boliver and Rodriguez from E.coli.
• Size is 4.3kbp
• Constructed from the pBR 318, pBR320.
• It contain Origin of replication.
4. • It also contain antibiotic resistant gene i.e. ampicillin
and tetracycline resistant gene .
• In ampicillin region it contain restriction site for
enzyme Pvu I and Pst I.
• In tetracycline region it contain restriction site for
enzyme Bam HI, Sal I.
5. ii) pUC19 Vector
• Messing and coworker developed at university of
California hence named pUC.
•Size is 2700 bp( 2-7 kbp)
•It possess lacZ gene as a marker gene and ori Region,
•It possess ampicillin resistant gene encoding restriction
site of enzyme Xmn L and Sca L.
6. Cosmid as a vector
• Hybrid vector derived from plasmid contain cos site
of phage λ
•Developed by Collins and Hoan in 1978
•It contains a ColE1 origin of replication
•Size is 5 Kbp
•The cod site is 12bp
7. Phage as a vector
•Bacteriophage are the viruses that infect bacterial cell.
•The viral DNA replicates and express inside the
bacterial cell and produces number of phage particles.
•The ability of transforming the viral DNA from the
capsid of phage design them as a vector.
• e.g.M13 , λ
Phage λ 10
• Phage λ 10 is 45 Kbp.
•Double stranded DNA for cloning is only 7 kbp in
length
•The cos site is the key feature.