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首页> 外文期刊>Cytology and genetics >Transformation and evaluation of different transgenic lines for Glyphosate tolerance and cane borer resistance genes in sugarcane (Saccharum officinarum L.)
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Transformation and evaluation of different transgenic lines for Glyphosate tolerance and cane borer resistance genes in sugarcane (Saccharum officinarum L.)

机译：不同转基因品系对甘草（Saccharum officinarum L.）的草甘膦耐受性和甘蔗bore虫抗性基因的评估

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摘要

The aim of this study was to ensure the systematic protein expression of two genes (GTG and Cry1Ac) under the influence of two different constitutive promoters i.e. Ubiquitin-1 and CaMV 35S promoters in different sugarcane lines. PCR amplification of GTG and Cry1Ac was achieved from putative transgenic plants through gene specific primers. Qualitative comparisons of GTG and Cry1Ac genes expression under two different promoters were obtained through protein dot blot and dipstick assay. The appearance of comparatively dark color dots in dot blot and dark color bands on dipstick with Ubiquitin as compared to light color bands with CaMV35S promoter, qualitatively confirmed high protein expression of two genes under Ubiquitin promoter. In quantitative gene expression comparisons maximum optical density (OD) at 450 nm of UV-light was obtained for GTG (3.7 OD) and Cry1Ac (3 OD) under Ubiquitin promoter, while for GTG (1.6 OD) and Cry1Ac (2.5 OD) with CaMV 35S promoter. The results indicated higher expression of two genes under Ubiquitin-1 promoter in sugarcane was found as compared to CaMV 35S promoter. This study provides a guide for stable and high expression of transgenes with reference to Ubiquitin-1 promoter which can be utilize in sugarcane as well as in other monocots.
机译：这项研究的目的是确保在两个不同的组成型启动子即Ubiquitin-1和CaMV 35S启动子在不同甘蔗品系的影响下，两个基因（GTG和Cry1Ac）的系统蛋白表达。通过基因特异性引物从推定的转基因植物中获得了GTG和Cry1Ac的PCR扩增。通过蛋白质斑点印迹和量油尺测定获得了两个不同启动子下的GTG和Cry1Ac基因表达的定性比较。与具有CaMV35S启动子的浅色带相比，在泛素的斑点印迹中出现了较暗的色点，在试纸上出现了深色带，定性地证实了泛素启动子下两个基因的高蛋白表达。在定量基因表达比较中，泛素启动子下的GTG（3.7 OD）和Cry1Ac（3 OD）在450 nm紫外光下获得了最大光密度（OD），而GTG（1.6 OD）和Cry1Ac（2.5 OD）在紫外光下获得了最大光密度（OD）。 CaMV 35S启动子。结果表明，与CaMV 35S启动子相比，在甘蔗中Ubiquitin-1启动子下两个基因的表达更高。这项研究提供了有关泛素-1启动子的稳定和高表达转基因的指南，该启动子可用于甘蔗以及其他单子叶植物。

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