Evaluation Of Fusion Peptides As Novel Gene Carriers Into Nicotiana Benthamiana And Arabidopsis Thaliana Leaves

• Main Author: Lakshmanan, Manoj Kumar
• Format: Thesis
• Language: English
• Published: 2016
• Subjects: QH1 Natural history (General - Including nature conservation, geographical distribution)
• Online Access: http://eprints.usm.my/31372/1/MANOJ_KUMAR_LAKSHMANAN_24.pdf; http://eprints.usm.my/31372/

Penghantaran gen menggunakan peptida melibatkan proses pengenalan gen asing ke dalam sel haiwan dan tumbuhan melalui peptida berfungsi yang mengandungi domain penembus sel dan jujukan polikation. Peptida penembus sel berkebolehan untuk memasuki sel-sel hidup, sementara jujukan polikation berkebolehan untuk bercantum dan memadatkan plasmid DNA secara elektrostatik. Dalam kajian ini, potensi peptida gabungan yang melingkungi unjuran polikation dan domain berfungsi dengan berkebolehan untuk menembusi sel hidup telah dibentuk dan diuji sebagai pengangkut gen ke dalam sel tumbuhan. Peptida-peptida gabungan yang mempunyai unjuran polikation, nona-arginin (R9) dan ko-polimer histidin-lisina (KH)9 berserta peptida penembus sel Bp100 (KKLFKKILKYL) dan Tat2 (RKKRRQRRRRKKRRQRRR) telah dicampurkan dengan plasmid DNA (pDNA) yang mengekod Renilla luciferase (RLuc) untuk membentuk kompleks pDNA-peptida pada pelbagai nisbah N/P. Nisbah, N/P merujuk kepada nisbah bilangan kumpulan amina daripada peptide per bilangan kumpulan fosfat daripada pDNA. Peptide-based gene delivery involves introduction of foreign genes into animal and plant cells via functional peptides, containing cell penetrating domains and polycationic sequences. Cell penetrating domains are capable of being internalized into living cells, while polycationic sequences can electrostatically interact and condense DNA. In this study, three fusion peptides consisting of polycationic sequences and functional domains with cell penetrating ability were designed and evaluated as potential gene carriers for plant cells. The fusion peptides, consisting of nona-arginine (R9) and histidine-lysine (KH)9 polycationic sequences as well as Bp100 (KKLFKKILKYL) and Tat2 (RKKRRQRRRRKKRRQRRR) cell penetrating domains respectively were mixed with pDNA encoding Renilla luciferase (RLuc) to form pDNA-peptide complexes at various N/P ratios. Here, N/P ratio refers to the ratio of number of amines from peptides per number of phosphates from pDNA.