pUC 19 vector ( Microbial biotechnology)pptx

Microbial biotechnology

1. VIVEKANANDHA ARTS AND SCIENCE COLLEGE FOR WOMEN Veerachipalayam – 637303, Sankagiri, Salem Dt., Tamilnadu, India . Affiliated to Periyar University , Salem; Recognised Under section 2 (f)& 12(B) of the UGC Act,1956. SUBJECT INCHARGE: Dr.R. PANDIYAN , ASSISTANT PROFESSOR , DEPARTMENT OF MICROBIOLOGY , VIVEKANANDHA ARTS AND SCIENCE COLLEGE FOR WOMEN SANKAGIRI, SALEM. SUBMITTED BY: A.VIDHYA , III- B.SC (MICROBIOLOGY), DEPARTMENT OF MICROBIOLOGY, VIVEKANANDHA ARTS AND SCIENCE COLLEGE FOR WOMEN, SANKAGIRI, SALEM. DEPARTMENT OF MICROBIOLOGY SUBJECT: MICROBIAL BIOTECHNOLOGY TOPIC : pUC 19

2. pUC 19

3. SYNOPSIS Introduction pUC vector Structure Sites Applications Advantages Disadvantages

4. INTRODUCTION pUC plasmids were first developed by Joachim Messing and his co-workers. It is a commonly used cloning vector in the bacteria E. Coli.
pUC19 is 2686 bp in length. The molecular weight of the pUC19 vector is 1.75×106 Da. It is a small plasmid with a high copy number. It contains the lacz gene and has multiple cloning sites.
Hence, it is widely used as a cloning vector. pUC19 plasmid is similar to pBR322 plasmid in structure.

5. pUC VECTOR pUC vector is a plasmid used for cloning in bacteria, such as E. Coli. It is a small, double-stranded circle of DNA that has several advantages over other plasmids, including: High copy number Multiple cloning sites Ampicillin resistance Blue-white screening

6. HIGH COPY NUMBER pUC vector can have 500–600 copies per cell. MULTIPLE CLONING SITES The pUC vector has a polylinker with unique sites for 13 different restriction endonucleases.

7. Ampicillin resistance The pUC vector contains an ampicillin resistance gene, which makes it easy to select for the plasmid in E. Coli. Blue – white screening The pUC vector contains the lacz’ gene, which allows for blue-white screening on media containing IPTG and X-gal.

8. STRUCTURE

9. APPLICATIONS Cloning: The pUC19 vector can be used to clone and express genes of interest in E. Coli.
Blue-white selection: The pUC19 vector’s multiple cloning site (MCS) is inserted into the LacZ-gene, which allows for blue-white selection of plasmid DNA that contains inserts.
Amplifying cDNA sequences: The pUC19 vector can be used to amplify ORF cDNA sequences by PCR.

10. Subcloning: The pUC19 vector can be used for subcloning with gene specific primers.
DNA sequencing: The pUC19 vector is suitable for DNA sequencing by the dideoxy method.
Model plasmid: The pUC19 vector is frequently used as a model plasmid in research.

11. ADVANTAGES Small size: The small size of the pUC19 vector makes it easy to clone.
High copy number: The pUC19 vector has a high copy number, which makes it a preferred choice for molecular techniques.
Multiple cloning sites: The pUC19 vector has a multiple cloning site (MCS) with 13 unique restriction enzyme sites.
Ampicillin resistance: The pUC19 vector is ampicillin resistant.
Blue/white screening: The pUC19 vector can be used for blue/white colony screening.

12. Compatible with many competent cells: The pUC19 vector is compatible with most commercially available competent cells, such as TOP10, DH5α, TOP10F´, and JM109.
DNA sequencing: The pUC19 vector is suitable for DNA sequencing by the dideoxy method.
Cloning of DNA fragments: The pUC19 vector may allow successful cloning of DNA fragments longer than the M13 phage vector
Expression of cloned DNA: The pUC19 vector can be used to express cloned DNA by using the lac promoter.

13. DISADVANTAGES One disadvantage of the pUC19 vector is that it can’t accommodate genes of interest that are larger than 15 kilobases (kb).
A vector is a substance, usually a piece of DNA, that carries genetic material and introduces it into a new cell. Vectors are used in molecular cloning to introduce DNA inserts into host cells.