Transformasi Genetik Kedelai dengan Gen Proteinase Inhibitor II Menggunakan Teknik Penembakan Partikel

Abstract

<p class="p1">An experiment was conducted at the Molecular Biology and Genetic Engineering Laboratory of BB-Biogen, Bogor with an objective to obtain transgenic soybean plants containing the <em>proteinase inhibitor </em>II (<em>pin</em>II) gene. The experiment consisted of three steps, i.e., optimalization of the soybean transformation technique using the <em>gus </em>gene; transformation of soybean using the <em>pin</em>II gene, and molecular analysis of the transformed soybean plants. Two type of explants (young embryo and cotyledon) were bombarded with <em>p</em>RQ6 plasmid containing the <em>gus </em>gene with the following treatment: Helium gas pressure (1100 psi and 1300 psi), shoot distance (5 and 7 cm), and number of bombardment (1x and 2x). The result of <em>gus </em>assay indicated that the best bombardment was done on young cotyledon explants with 1100 psi Helium pressure, shoot distance 5 cm, and 1x bombardment. Transformation of the soybean explant using the <em>pin</em>II gene (inside the <em>p</em>TWa plasmid) was conducted using the best bombardment treatment from the first activity. Two plants from c.v. Wilis (WP<span class="s1">1</span>, WP<span class="s1">2</span>) and three plants from c.v. Tidar (TP<span class="s1">1</span>, TP<span class="s1">2</span>, TP<span class="s1">3</span>) were recovered from regeneration and selection of the transformed explants. Molecular analysis of the regenerated plants using the PCR technique showed that only WP<span class="s1">2 </span>contained the <em>pin</em>II gene. This plant was fertile and will be used for further evaluation.</p>