کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5591066 | 1570335 | 2016 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A super twin T-DNA vector that allows independent gene expression during Agrobacterium-mediated transformation
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کلمات کلیدی
GUSHYGMurashige and Skoog - Murashige و SkoogSouthern blot - بلوط جنوبCotransformation - تعاونیTobacco - توتون و تنباکوmg/L - میلی گرم / لیترpolymerase chain reaction - واکنش زنجیره ای پلیمرازPCR - واکنش زنجیرهٔ پلیمرازβ-glucuronidase gene - ژن β-گلوکورونیدازhygromycin resistance gene - ژن مقاومت هیگرموسین
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
ژنتیک
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
In this study, we designed and constructed a super twin T-DNA vector (pTRIDT313-g) containing two independent T-DNA cassettesâone for the selection gene Hyg and the other for the target gene Gusâto produce marker-free transgenic lines. The resulting vector was transformed into tobacco, and polymerase chain reaction (PCR) analysis showed four types of gene combinations in the T1 and T2 generations: Gus only, Hyg only, Gus + Hyg, and untransformed lines. The intermediate region from the T-DNA of the right border of Hyg to the left border of Gus in the Hyg and Gus lines was not amplified. Genome walking confirmed that the Hyg and Gus T-DNA cassettes were independently inserted in different regions of the tobacco genome. Thus, the two T-DNA cassettes were integrated randomly as independent loci into the tobacco genome. The results of reverse transcription-PCR indicated that Hyg could normally be expressed in the roots, stems, and leaves of transgenic lines, and the resistance test showed that all Hyg transgenic lines could grow in the presence of 50 mg/L hygromycin. All Gus transgenic lines showed obvious blue coloration in enzyme activity tests, indicating that the Gus gene could be normally expressed in all the lines. Therefore, the super twin T-DNA vector (pTRIDT313-g) exhibits independent integration, heredity, and normal gene function from two T-DNA cassettes. This vector could be a useful and valuable tool in the production of marker-free transgenic lines.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Plasmid - Volumes 87â88, SeptemberâNovember 2016, Pages 58-64
Journal: Plasmid - Volumes 87â88, SeptemberâNovember 2016, Pages 58-64
نویسندگان
Qiang Yang, Mei Deng, Ling-Ling Zhang, Xiao-Wei Zhang, Le-Ning Wang, Hu Chen, Jian Ma, Peng-Fei Qi, Qian-Tao Jiang, Xiu-Jin Lan, Yu-Ming Wei, You-Liang Zheng,