Utami Edy Setiti Wida, Hariyanto Sucipto, Manuhara Yosephine Sri Wulan
Laboratory of Plant Tissue Culture, Department of Biology, Faculty of Sciences and Technology, Universitas Airlangga, Surabaya, Indonesia.
Laboratory of Ecology, Department of Biology, Faculty of Sciences and Technology, Universitas Airlangga, Surabaya, Indonesia.
J Genet Eng Biotechnol. 2018 Dec;16(2):703-709. doi: 10.1016/j.jgeb.2018.02.002. Epub 2018 Feb 22.
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.
首次开发了一种由[具体介导方式]介导的遗传转化方案并生产了转基因[具体生物]。将8周龄的原球茎外植体用作转化靶点，用携带质粒pG35SKNAT1的[具体菌株]LBA4404进行转化。对感染时间、[具体细菌]密度、乙酰丁香酮浓度和共培养时间等几个参数进行了转化效率评估。数据采用单因素方差分析（ANOVA）和邓肯多重范围检验（DMRT）进行分析，p<0.05。随后，通过聚合酶链反应（PCR）分析确认了KNAT1基因的表达。从感染[具体细菌]培养物的原球茎外植体获得的最高转化效率（70%）是在OD浓度为0.6、感染30分钟，并与100µM乙酰丁香酮共培养5天时。PCR分析的确认结果表明，KNAT1基因已整合并在转基因[具体生物]的基因组中表达。
