TY - JOUR AU - handayani, Tri AU - Alimuddin, Alimuddin AU - Widyastuti, Utut AU - Suryati, Emma AU - Parenrengi, Andi PY - 2014/11/28 Y2 - 2024/03/29 TI - Binary Vector Construction and Agrobacterium tumefaciens-mediated Transformation of Lysozyme Gene in Seaweed Kappaphycus alvarezii JF - BIOTROPIA JA - BIOTROPIA VL - 21 IS - 2 SE - Research Paper DO - 10.11598/btb.2014.21.2.348 UR - https://journal.biotrop.org/index.php/biotropia/article/view/348 SP - 82-92 AB - <span style="font-size: 12.0pt; line-height: 115%;" lang="EN-US">Ice-ice disease is the biggest problem in the cultivation of seaweed <em>Kappaphycus alvarezii</em>. The disease is caused by bacterial infection and induced by drastic changes of water quality. <em>Lysozyme</em> has the ability to break down bacterial cell wall. The purpose of this research was to construct of a binary vector pMSH1-Lys carrying chicken <em>Lysozyme</em> (<em>Lis</em>) gene and introduce pMSH1-Lis on <em>K. alvarezii</em>. The binary vector expression was transformed into <em>Agrobacterium tumefaciens</em> LBA4404 by triparental mating. Thallus was inoculated with <em>A. tumefaciens</em> carrying pMSH1-Lis and then the transformed thallus was selected by adding 20 mg/L hygromycin to the culture medium. PCR analysis showed that the construction of the binary plasmid pMSH1-Lys was established. Percentage of transformation of pMSH1-Lis on <em>K. alvarezii</em> was 23.56%, while the efficiency of regeneration was 11.32%. PCR analysis showed that three of the regenerated thallus contained <em>Lysozyme</em> gene. Thus, transgenic <em>K. alvarezii</em> was produced successfully and this can be useful for studying the mechanisms of seaweed defense against bacterial infection.</span> ER -