Zagazig University Digital Repository
Home
Thesis & Publications
All Contents
Publications
Thesis
Graduation Projects
Research Area
Research Area Reports
Search by Research Area
Universities Thesis
ACADEMIC Links
ACADEMIC RESEARCH
Zagazig University Authors
Africa Research Statistics
Google Scholar
Research Gate
Researcher ID
CrossRef
The pJan25 vector series: An enhancement of the Gateway-compatible vector pGWB533 for broader promoter testing applications
Faculty
Agriculture
Year:
2013
Type of Publication:
Article
Pages:
249-256
Authors:
Alzohairy, Ahmed Mansour, MacDonald, Margaret H, Matthews, Benjamin F
DOI:
10.1016/j.plasmid.2013.01.005
Journal:
PLASMID ACADEMIC PRESS INC ELSEVIER SCIENCE
Volume:
69
Research Area:
Genetics \& Heredity
ISSN
ISI:000319368500007
Keywords :
Plasmid, pJan25, Gateway, eGFP, GUS, Promoter
Abstract:
Agrobacterium-mediated transformation of plants has enhanced our ability to progress more rapidly in plant genetic engineering. Development of binary vectors for Agrobacterium has played a major role in advancing plant biology. Here, we report new features added to the Gateway-compatible vector pGWB533 for promoter testing with the reporter gene encoding beta-glucuronidase (GUS). The original vector contains the spectinomycin/streptomycin adenylyltransferase (aadA) gene for bacterial selection and the hygromycin phosphotransferase gene (hpt) for transformed plant selection. However, some bacterial strains used to transform plants, such as Agrobacterium rhizogenes strain K599, have elevated tolerance to spectinomycin and streptomycin, thus making bacterial selection of pGWB533 inefficient. Although pGWB533 confers chemical selection for transgenic plants using hygromycin resistance, the plasmid has no visual marker that enables visual selection of transformed plants or transgenic tissue. In this regard, adding a gene to constitutively express green fluorescent protein (eGFP) makes it easier to visually select the transformed tissue and trim out the non-transformed. In this report we describe a series of vectors, pJan25S (NCBI: KC416200), pJan25T (NCBI: KC416201) and pJan25X (NCBI: KC416202), that are enhancements of pGWB533 for promoter testing. All three vectors contain the gene encoding eGFP as a visual marker for transformed tissue. However, in pJan25S and pJan25T, eGFP is controlled by the rolD promoter for root-specific expression, while in pJan25X it is controlled by the CaMV35S promoter for constitutive expression in all plant tissues. Spectinomycin and streptomycin resistance remains in pJan25S for bacterial selection; however, pJan25T and pJan25X contain the gene encoding tetracycline resistance (tet) for bacterial selection. These changes resulted in enhanced vectors with better visual and chemical selection that should have broad application in promoter studies. Published by Elsevier Inc.
Online
PDF
جامعة المنصورة
جامعة الاسكندرية
جامعة القاهرة
جامعة سوهاج
جامعة الفيوم
جامعة بنها
جامعة دمياط
جامعة بورسعيد
جامعة حلوان
جامعة السويس
شراقوة
جامعة المنيا
جامعة دمنهور
جامعة المنوفية
جامعة أسوان
جامعة جنوب الوادى
جامعة قناة السويس
جامعة عين شمس
جامعة أسيوط
جامعة كفر الشيخ
جامعة السادات
جامعة طنطا
جامعة بنى سويف