Development ofselectable marker free,insect-resistant,transgenic mustard (Brassica juncea)plants usingCre/lox mediated recombination

1. ZEESHAN NADEEM
ROLL NO. 26
BS. BIOTECHNOLOGY 7TH
IBB, PU

2. DEVELOPMENT OF
SELECTABLE MARKER FREE,
INSECT-RESISTANT,
TRANSGENIC MUSTARD (Brassica juncea)
PLANTS USING
CRE/LOX MEDIATED RECOMBINATION
BY
ARPITA BALA, AMIT ROY, AYAN DAS,
DIPANKAR CHAKRABORTI AND SAMPA DAS

3. INTRODUCTION
» Brassica juncea (Indian Mustard) an important crop
» Lipaphis erysimi (a sap sucking hemipteran pest)

4. OBJECTIVES
» Development of Insect resistance by:
Expressing Allium sativum leaf agglutinin (ASAL)
gene (replacement of chemical pesticides and cry toxins)
» Removal of Selectable Marker Genes (hpt) by:
Using Cre/lox mediated recombinantion

5. CRE LOX MEDIATED RECOMBINATION

6. VECTOR CONSTRUCTS

7. RESULTS…

8. Southern Blot Analysis of Expression of ASAL
and Cre Genes

9. Multiplex PCR Analysis of Marker Gene
Excision

10. Western Blot Analysis of ASAL Expression in
F1 Hybrid Plants

11. Agglutination and Thermal Stability Assay of
Expressed ASAL Protein from F1 Hybrids

12. In-Planta Insect Bioassay
Insect Bioassay Setup

14. PCR Analysis of Completely Marker Free F2
Mustard Plants

15. CONCLUSION
Efficient expression of insect resistant ASAL gene
in Brassica juncea plants
even after the elimination of selectable marker gene
by Cre/lox mediated recombination