Synthetic Coloning Vector Type (BAC)

1. BACTERIAL
ARTIFICIAL
CHROMOSOME
AFNAN MALIK, ALEENA KHALID, SHOAIB IQBAL,
SARMAD DURRANI

2. INTRODUCTIO
N
Natural
chromosome:
Thread like
structure of DNA
and protein.
Properties of
Artificial
Chromosome.
• MCS
• ORI
• Selectable Marker
Artificial
chromosome.
 BAC
 YAC
 MAC

3. are specialized DNA
vectors derived from the
naturally occurring F
(fertility) plasmid
in Escherichia coli (E. coli).
BAC
Nucleus
First artificial
chromosome was
designed by Hiroaki
Shizuya in 1992 at
California Institute of
Technology by altering
plasmid F factor.
HISTORY
SIZE: 100-300KB

4. • Origin of Replication
• Par A and B
• Selectable marker
• T7 and Sp6
Component
s

5. • CMR is a selectable
marker
• Ori E, repE, par A and
b
• Hind III and BamHI
• NotI as cutting out
inserted fragment
Structure of
BAC

6. How can we
make?
• Insert preparation
• Ligation
• Transformation
• Selection
• Propagation
• Analysis

7. BAC VS YAC
Feature BACs YACs
Host Organism Escherichia coli (bacteria) Saccharomyces cerevisiae (yeast
Insert Size 100–300 kilobases (kb) 100–2000 kilobases (kb)
Stability
High (low-copy replication,
minimal rearrangements)
Low (prone to recombination
and instability)
Ease of Use Easy to manipulate in E. coli
Complex (requires yeast
handling and specialized
techniques)
Copy Number Low (1–2 copies per cell) Low (1–2 copies per cell)
Structural Components
Derived from the F plasmid;
includes oriS, par genes, and
selectable markers
Contains yeast centromere,
telomeres, and autonomous
replication sequence (ARS)
Applications
Genomic libraries, genome
sequencing, functional
studies
Large genome mapping,
cloning of extremely large DNA
fragments

8. ROLE IN RECOMBINANT
PROTEIN TECHNOLOGY
• High Cloning Capacity
• Stability in Host Cells
• Ease of Manipulation
• Low-Copy Replication
• Faithful Gene Expression
• Reduced Risk of Silencing
• Applications in Mammalian Cell
Expression Systems
• Functional Genomics
• Recombinant Protein Production
• Gene Therapy and Vaccine
Development

9. Advantages and Limitation
Advantages
• High stability
• Very efficient
• Large carrying capacity
• Low rate of chimerism
• Ease with handling Limitation
• Lacks positive selection
for clones
• Very low yield

10. • Researchers are investigating herpes virus, used BAC vector
that are cultured in bacterial cells and when transferred in
animal cell, release inserted DNA.
• It has also impact on phylogenetic studies (i.e relationship
of species to one another). In present era we have
abundance of microbial species all over around the world
which cannot be grown on cultures.
• BACs are also in use to study pathogens and in designing
new vaccines.
• Modification of modal organism.
Applications

11. Case study
• Source DNA preparation
• Vector Preparation
• Ligation
• Selection and screening
• Characterization

12. Advances
• Enhanced cloning capacity
• Improved stability and fidelity
• Automation and HTS
• CRISPR CAS9 System
Improvement
• Optimized regulatory elements
• Epigenetic modification
• Host cell engineering
• Inducible expression system

13. THANKS
Question and Answers
The end of the
presentation