Finally

MolecularCloning.com

Phew... Finally I managed to pull out everything I got to make my thesis.. Here's my Introduction:

Genetic engineering of plants has become an important tool for crop improvement. Through this technique, it is now possible to create transgenic plants that are resistant to insects and pathogens or adapted to specific soil or environmental conditions. However, engineering of transgenic resistance in plants requires suitable components and strategy, so that maximum performance may be achieved. Such strategy and components are lacking when come to engineering resistance in local crops.

To develop transgenic resistance in plants against major pathogens requires proper promoter system in regulating expression genes in plants. The purpose of this study is to develop proper promoter system to regulate plant expression gene constructs in plants. A potential promoter candidate is Phenylalanine ammonia-lyase (PAL) gene promoter known to be involved in lignification process.

In this project, a phenylalanine ammonia-lyase (PAL) gene fragment will be amplified, cloned and sequenced from Musa acuminata cv Jaribuaya. The PAL sequence will be then compared with the sequences of PAL genes available in online databases.

Therefore objectives of this research were:
To amplify PAL gene fragment from Musa acuminata cv. Jari buaya (banana).
To clone and sequence the PAL gene fragment.
To analyze the sequence of the cloned PAL gene fragment by BLAST analysis.

and Here's my Abstract:
Genetic engineering of plants has become an important tool for crop improvement. To develop transgenic resistance in plants against major pathogens requires proper promoter system in regulating expression genes in plants. The enzyme phenylalanine ammonia-lyase (PAL) was found to be encoded by a small gene family in the Musa acuminata cv Jari buaya. At least five of the family members are tightly clustered within approximately 700bp of DNA. Amplification of PAL gene by PCR using degenerate primers PALFor1EcoR1 (5’ – GAAGATCTTCGVTTCTTGAATGABGGAATYTTTYGGNA – 3’), PALFor2EcoR1
(5’ – GAAGATCTTCAACCAAGATGTBAACTCNYNGGGNT – 3’), PALFor3EcoR1
(5’ – GAAGATCTTCTBCTCCAGGNTACTCNGGNATCAGA – 3’), and PALRevBgIII (5’ – CGGAATTCCGTTADCAHATNGGAAGAGGAGCACCA – 3’) produced multiple bands of four. A 350bp PAL gene fragment was cloned into the pGEM-T vector and transformed into competent cells, E.coli JM109. The recombinant plasmid, pGEM-PAL was isolated from the bacterial cells and digested with SalI producing a fragment of 3350bp nucleotides, a linearized plasmid. The sequencing however yields a 150bp PAL gene fragment cloned. BLAST analysis showed the sequence is not similar with any other organisms PAL gene fragment.


Tell me what do you guys think...