-mediated transformation of J.J.Sm: An important medicinal orchid.

-mediated transformation of J.J.Sm: An important medicinal orchid.

Utami, Edy Setiti Wida;Hariyanto, Sucipto;Manuhara, Yosephine Sri Wulan;
journal, genetic engineering & biotechnology 2018 Vol. 16 pp. 703-709
157
utami2018mediatedjournal

Abstract

A protocol for genetic transformation mediated by and production of transgenic has been developed for the first time. The 8-week-old protocorm explants were used as target of transformation with strain LBA4404 carrying plasmid pG35SKNAT1. Several parameters such as infection period, density, concentration of acetosyringone, and co-cultivation period were evaluated for the transformation efficiency. The data were analyzed using one-way analysis of variance (ANOVA) and Duncan's Multiple Range Test (DMRT) with p < 0.05. Subsequently, KNAT1 gene expression was confirmed by polymerase chain reaction (PCR) analysis. The highest efficiency of transformation (70%) obtained from protocorm explants infected with culture was at the OD concentration of 0.6 for 30 min, and co-cultivated with acetosyringone 100 µM for 5 days. The results of confirmation by PCR analysis show that the KNAT1 gene has been integrated and expressed in the genome of transgenic.

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0x95644003c57E6F55A65596E3D9Eac6813e3566dA
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32792
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10.1016/j.jgeb.2018.02.002
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