Salient features of cloning vector

1. PLASMID AS CLONING
VECTOR
Jean de Dieu BAZIKI
PhD Scholar

2. OUTLINE
• Introduction
• Features
• Types
• Advantages
• Disadvantages
• References

3. Objective
 Describe salient features for ideal
plasmid as cloning vector

4. Introduction
 Plasmid
Extrachromosal
Self replicating
Short double stranded
Closed Circular DNA molecules
Found in prokaryotes
 Small size

5. Types of plasmids by their
Functions
 Col plasmids
 Fertility-F plasmids
 Resistance plasmids
 Degradative plasmids
 Virulence plasmids
 Others
Ti plasmids
Ri plasmids

6. Plasmids as cloning vectors
 In 1973, Chang and Cohen proved the
use of plasmid as cloning vector in
genetic engineering/ Biotechnology .

7. Ideal features plasmid as cloning
vector
 Small size 1.2-3kb
 Origin of replication
 Unique restriction site
 Multiple cloning site
 Selection marker
 Copy number
 Control elements ( Promoter, operator,
ribosomal binding sites)
 No pathogenicity

10. 3 important features: Cloning site, Ori-an origin
of replication, A selectable marker (ampr)

13. Advantages
• Small, easy to handle
• Straightforward selection
strategies
• Useful for cloning small
DNA fragments(< 10kbp)

14. Disadvantages
• Less useful for cloning
large DNA fragments (>
10kbp)

15. Application
 A particular gene can be isolated and its
nucleotide sequence be determined
 Genomic library construction
 Preparation of probes
 Control sequences DNA can be identified and
analysed
 Protein/enzyme/RNA function can be
investigated
 Mutations can be identified e.g: gene defects
related to specific diseases
 Organism can be “engineered” for specific
purposes e.g: Insulin production

16. References
Cohen, S. N., Chang, A. C., Boyer, H. W., & Helling, R. B. (1973). Construction of
biologically functional bacterial plasmids in vitro. Proceedings of the National Academy
of Sciences of the United States of America, 70(11), 3240–4. Retrieved from
http://www.ncbi.nlm.nih.gov/pubmed/4594039
full-text. (n.d.).
Gene Cloning and DNA Analysis: An Introduction - T. A. Brown - Google Books. (n.d.).
Retrieved February 24, 2017, from
https://books.google.co.ke/books?hl=en&lr=&id=pi3dCQAAQBAJ&oi=fnd&pg=PR17
&dq=plasmid+cloning+vector+features&ots=C3_dR165S5&sig=LM0J_IErGQcSujVDa
KydASzErg8&redir_esc=y#v=onepage&q=plasmid cloning vector features&f=false
Promega. (2010). pGEM®- T and pGEM®- T Easy Vector Systems. Technical Manual, 1–
28.

17. THANK YOU
MERCI
MURAKOZE (Rwanda)
ASANTE SANA (Kenya)
NAMASIGNALO (Ethiopia)
YEKENIYELLEY (Eritrea)
GANGI DEBE (Chad)
MASIAKNEE (Cameroon)
WEBALE (Uganda)