Construction of plant expression vector harbouring Oak 1 gene encoding kalata B1 protein

2019 ◽  
Author(s):  
Norsharina Md Saad ◽  
Teo Chee How ◽  
Zuraida Ab Rahman ◽  
Zamri Zainal
2004 ◽  
Vol 1 (2) ◽  
pp. 85-91 ◽  
Author(s):  
Wang Xian-Yan ◽  
Shan Xiao-Yi ◽  
Yang Ai-Fang ◽  
Zhang Ju-Ren

AbstractThe plant expression vector pCAMBIA1300-AtNHX1-als was constructed by inserting the herbicide resistance gene als of Arabidopsis thaliana into the plasmid pCAMBIA1300-AtNHX1, which contains the AtNHX1 gene encoding the Na+/H+ antiport from the vacuolar membrane of A. thaliana. Transgenic tobacco plants were obtained via Agrobacterium-mediated transformation. PCR and Southern blot assay indicated that genes als and AtNHX1 had been integrated into the genome of the transgenic plants. The herbicide resistance and salt tolerance of transgenic plants increased by about 1000-fold and by 1.5% NaCl concentration, respectively, compared with controls. Herbicide resistance of the T1 progeny was evaluated by spraying transgenic plants with different concentrations of Luhuanglong at the four-leaf stage. Controls gradually died under 100 mg/l Luhuanglong whereas 73% of the T1 plants still survived at 500 mg/l Luhuanglong. Thus the plant expression vector pCAMBIA1300-AtNHX1-als could be used to improve the herbicide resistance and salt tolerance of crops.


2020 ◽  
Vol 28 (S2) ◽  
Author(s):  
Fauziah Abu Bakar ◽  
Pavitra Paramalingam ◽  
Kamariah Hasan

Carica papaya is a well-liked and economically important fruit with outstanding nutritional and medicinal values. Its susceptibility to abiotic stress which affects the growth and harvest, causes significant yield loss to farmers. In recent years, significant progress has been made to understand the genes that play critical roles in abiotic stress response, especially some transcription factor (TF) encoding genes. Among all TFs, WRKY TF gene family is one of the best-studied TFs involved in various stress responses. To date, only limited information on functionally characterised WRKY TFs is available for C. papaya. The aim of this study was to produce a recombinant construct harbouring WRKY gene in pGEM®-T Easy cloning vector. The presence of a DNA band of the expected size of 465 bp on agarose gel electrophoresis indicated that WRKY gene was successfully amplified from all treated samples. DNA sequencing analysis revealed that the amplified sequence isolated from the treated samples were closely related to Carica papaya species with 97% similarity. Following transformation, 4 out of 5 colonies that were randomly selected showed the WRKY gene had been successfully inserted into pGEM®-T Easy vector and transformed into E. coli. In future, the WRKY gene from pGEMT-WRKY recombinant construct will be cloned into the plant expression vector pCAMBIA 1304 prior to transformation in the plant. The success of demonstrating the WRKY gene towards the response in abiotic stress will enable us to produce stress tolerant transgenic crops under unfavourable conditions via genetic engineering for sustained growth.


2009 ◽  
Vol 3 (1) ◽  
pp. 55-59 ◽  
Author(s):  
Chunli Song ◽  
Junlian Ma ◽  
Xia Tang ◽  
Zide Zhang ◽  
Pingping Zhou ◽  
...  

2013 ◽  
Vol 690-693 ◽  
pp. 1314-1317
Author(s):  
Xiang Li Song ◽  
Zhi Hui Wu ◽  
Xi Zhang ◽  
Meng Xi Lu ◽  
Bei Guo

In order to transcription and expression an exogenous gene just in transgenic strawberry fruit, the gene encoding tomato fruit-specific E8 promoter was cloned. We constructed plant expression vector with E8 promoter replace the CaMV35S of pBI121 that used for transformation of strawberry. Recombination plasmid was identified by PCR, restriction enzymes digestion and sequencing analysis, and transfered into the agrobacterium EHA105 strain. At the same time, the agrobacterium EHA105, transformation and no transformation tissue of strawberry used as the experiment materials to study the expression of reporter genegusAby the histochemical staining method. The results show that agrobacterium which containsgusAgene and strawberry organization of transformation can be dyed blue, others were not stain. So in testing the transgenic plants by agroinfiltration transient expression system can reduce or avoid false positive appearance.


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