Sudhir Kumar's document discusses bacteriophage vectors used for cloning DNA fragments. It describes how lambda phage vectors can clone fragments up to 25 kb by removing the phage's non-essential region. Two main types of lambda vectors are insertion vectors like lambda gt10 that insert foreign DNA, and replacement vectors like lambda DASH that replace phage genes. The document also briefly explains the M13 bacteriophage genome and references a textbook on gene cloning.

1. SUDHIR KUMAR
B.Sc (ZBC)
DEPARTMENT OF MICROBIOLOGY
DR. SHAKUNTALA MISRA NATIONAL REHABILITATION UNIVERSITY, MOHAN ROAD
LUCKNOW
2017-2018
Vectors

2. INTRODUCTION
 Bacteriophage vectors are used to clone large pieces of DNA, from 5 to 25 kb.
Used in preparing genomic library.
If the total size of the molecule is more than 52kb then it cannot be package into
phage head.
Phage generally used in cloning( infects E.coli ):-
λ-phage.

3. λ-phage Vector
NON-ESSENTIAL REGION
(19-35)
Non-essential region of the λ genome contains genes involve in
integration and excision of λ prophage from E.coli chromosome.
 λ DNA has multiple recognition sites for almost all restriction
endonucleases.
Removal of non essential region blocks lysogenic cycle, and gene
of interest is ligated.

4. Types of λ-phage Vector
INSERTION VECTOR
Lambda gt10 Lambda ZAP||

5. REPLACEMENT VECTOR
Lambda DASH||

6. M-13 BACTERIOPHAGE GENOME
Contains intergenic sequences of 507 nucleotide.
These sequences contains ORI which should be kept intact during
insertion of polylinkers and removal of intergenic sequences.

7. CONSTRUCTION OF M13mp1

8. REFERENCES
Brown, T.A. (2016). Gene Cloning and DNA Analysis 7th Edition (pp. 99-108)
Manchester, U.K. WILEY Blackwell.