Expression vector p301-bG1 contains a Sw gene and a bialaphos resistance gene both driven by glyceraldehydes-3-phosphate dehydrogenase (GPD) gene promoter isolated from Lentinus edodes ( Berk.) Sing. Using p301-bG1, P...Expression vector p301-bG1 contains a Sw gene and a bialaphos resistance gene both driven by glyceraldehydes-3-phosphate dehydrogenase (GPD) gene promoter isolated from Lentinus edodes ( Berk.) Sing. Using p301-bG1, PEG-mediated transformation of protoplast of L. edodes was studied. Mixed with PEG-purified plasmid DNA, the protoplasts of L. edodes were treated with PEG solution and cultured on CYM regeneration plate containing 40 mug/mL bialaphos. Bialaphos-resistant and GUS-positive transformants were obtained using this transformation system. Although the transformation efficiency was relatively low, the protocols release large expenses on expensive instrument and restriction enzymes, providing a simple and economical method for mushroom breeding at the molecular level.展开更多
Tremellafuciformis is one of higher basidiomycetes. Its basidiospore can reproduce yeast-like conidia, which is also called the blastospore by budding. The yeast-like conidia of T. fuciformis is monokaryotic and easy ...Tremellafuciformis is one of higher basidiomycetes. Its basidiospore can reproduce yeast-like conidia, which is also called the blastospore by budding. The yeast-like conidia of T. fuciformis is monokaryotic and easy to culture by submerged fermentation similar to yeast. Thus, it is a good recipient cell for exogenous gene expression. In this study, the expression plasmid pAN7-1 （containing promoter gpd-An derived from Aspergillus nidulans and selectable marker gene hph conferring resistance to hygromycin B） and plasmid pLg-hph （containing promoter gpd-Le derived from Lentinula edodes and selectable marker gene hph） were transformed into the yeast-like conidia of T. fuciformis by PEG-mediated protoplast transformation, respectively. The primary putative transformants were selected by the sandwich screening method with the selective medium containing 50 μg mL^-1 hygromycin. The putative transformants were obtained from the primary putative transformants transferred on PDSA plates containing 100 μg mL^-1 hygromycin for second round selection. Experimental results showed that the optimal concentration of PEG 4000 for mediating protoplast transformation was 25%. PCR and Southern blotting confirmed that the selectable marker gene hph was integrated effectively into the genome of the yeast-like conidia of T. fuciformis with plasmid pLg-hph transformation. Its transformation efficiency was 110 transformants per μg DNA, and the hph gene was integrated into the genome of some yeast-like conidia with plasmid pAN7-1 transformation. However, its transformation efficiency was only 9 transformants per μg DNA. The presence of hph gene in the genome of transformants after 5 generations of sub- culturing on PDSB medium was confirmed by PCR, suggesting that the foreign gene hph was stable during subculture.展开更多
In this study, DERB1A transcription factor and stress-induced promoter rd29A were isolated respectively and amplified from Arabidopsis thaliana, se- quenced and analyzed by DNAsis. In addition, the stress-induced prom...In this study, DERB1A transcription factor and stress-induced promoter rd29A were isolated respectively and amplified from Arabidopsis thaliana, se- quenced and analyzed by DNAsis. In addition, the stress-induced promoter rd29A was utilized to construct the plant expression vector of DERB1A, which was transformed into Agrobacterium tumefaciens. Furthermore, the transgenic regeneration system of fresh-cut chrysanthemum from callus to plantlets was established successfully. On this basis, chrysanthemum leaf-disc explants were genetically transformed with Agrobacterium-mediated method. Two positive transgenie plantlets were obtained in vitro. Based on PCR detection, DREB1A transcription factor was integrated into chrysanthemum genome, which laid the foundation for breeding new transgenie cultivars of fresh-cut chrysanthemum with high comprehensive stress resistance, good cmalitv and high field.展开更多
The 14th Beijing International Automotive Exhibition(Auto China 2016)will take place from April 25 to May 4,2016.It will be at China International Exhibition Center New Venue(Tianzhu)and China International Exhibition...The 14th Beijing International Automotive Exhibition(Auto China 2016)will take place from April 25 to May 4,2016.It will be at China International Exhibition Center New Venue(Tianzhu)and China International Exhibition Center Old Venue(Jinganzhuang)simultaneously and the total exhibition area is expected to be 220,000 square meters.Auto China 2016 is sponsored by China Machinery Industry展开更多
“Yang promotes Qi transformation and Yin promotes forming”maintains the normal progress of human life activities,which is an important embodiment of human vitality.The dysfunction of“Yang promotes Qi transformation...“Yang promotes Qi transformation and Yin promotes forming”maintains the normal progress of human life activities,which is an important embodiment of human vitality.The dysfunction of“Yang promotes Qi transformation and Yin promotes forming”leads to the coagulation of pathological products such as phlegm and blood stasis in the breast meridian,resulting in the occurrence of acne mastitis.In clinical treatment,we should take“warm Yang to promote Qi transformation”as the treatment principle,and add or subtract according to the specific situation.展开更多
文摘Expression vector p301-bG1 contains a Sw gene and a bialaphos resistance gene both driven by glyceraldehydes-3-phosphate dehydrogenase (GPD) gene promoter isolated from Lentinus edodes ( Berk.) Sing. Using p301-bG1, PEG-mediated transformation of protoplast of L. edodes was studied. Mixed with PEG-purified plasmid DNA, the protoplasts of L. edodes were treated with PEG solution and cultured on CYM regeneration plate containing 40 mug/mL bialaphos. Bialaphos-resistant and GUS-positive transformants were obtained using this transformation system. Although the transformation efficiency was relatively low, the protocols release large expenses on expensive instrument and restriction enzymes, providing a simple and economical method for mushroom breeding at the molecular level.
基金funded by the National Hi-Tech Research and Development Program of China (863 Program,2006AA10Z301)the National Natural Science Foundation of China (30371000, 30671457)
文摘Tremellafuciformis is one of higher basidiomycetes. Its basidiospore can reproduce yeast-like conidia, which is also called the blastospore by budding. The yeast-like conidia of T. fuciformis is monokaryotic and easy to culture by submerged fermentation similar to yeast. Thus, it is a good recipient cell for exogenous gene expression. In this study, the expression plasmid pAN7-1 （containing promoter gpd-An derived from Aspergillus nidulans and selectable marker gene hph conferring resistance to hygromycin B） and plasmid pLg-hph （containing promoter gpd-Le derived from Lentinula edodes and selectable marker gene hph） were transformed into the yeast-like conidia of T. fuciformis by PEG-mediated protoplast transformation, respectively. The primary putative transformants were selected by the sandwich screening method with the selective medium containing 50 μg mL^-1 hygromycin. The putative transformants were obtained from the primary putative transformants transferred on PDSA plates containing 100 μg mL^-1 hygromycin for second round selection. Experimental results showed that the optimal concentration of PEG 4000 for mediating protoplast transformation was 25%. PCR and Southern blotting confirmed that the selectable marker gene hph was integrated effectively into the genome of the yeast-like conidia of T. fuciformis with plasmid pLg-hph transformation. Its transformation efficiency was 110 transformants per μg DNA, and the hph gene was integrated into the genome of some yeast-like conidia with plasmid pAN7-1 transformation. However, its transformation efficiency was only 9 transformants per μg DNA. The presence of hph gene in the genome of transformants after 5 generations of sub- culturing on PDSB medium was confirmed by PCR, suggesting that the foreign gene hph was stable during subculture.
基金Supported by Natural Science Foundation of Yunnan Province(2007C213M)Provincial Key Discipline of Landscape Plant&Ornamental Horticulture of Yunnan Province+1 种基金Provincial Key Lab of Colleges and Universities in Landscape Plants and Ornamental Horticulture of Yunnan ProvinceLarge Apparatuses Sharing Platform of Southwest Forestry University
文摘In this study, DERB1A transcription factor and stress-induced promoter rd29A were isolated respectively and amplified from Arabidopsis thaliana, se- quenced and analyzed by DNAsis. In addition, the stress-induced promoter rd29A was utilized to construct the plant expression vector of DERB1A, which was transformed into Agrobacterium tumefaciens. Furthermore, the transgenic regeneration system of fresh-cut chrysanthemum from callus to plantlets was established successfully. On this basis, chrysanthemum leaf-disc explants were genetically transformed with Agrobacterium-mediated method. Two positive transgenie plantlets were obtained in vitro. Based on PCR detection, DREB1A transcription factor was integrated into chrysanthemum genome, which laid the foundation for breeding new transgenie cultivars of fresh-cut chrysanthemum with high comprehensive stress resistance, good cmalitv and high field.
文摘The 14th Beijing International Automotive Exhibition(Auto China 2016)will take place from April 25 to May 4,2016.It will be at China International Exhibition Center New Venue(Tianzhu)and China International Exhibition Center Old Venue(Jinganzhuang)simultaneously and the total exhibition area is expected to be 220,000 square meters.Auto China 2016 is sponsored by China Machinery Industry
文摘“Yang promotes Qi transformation and Yin promotes forming”maintains the normal progress of human life activities,which is an important embodiment of human vitality.The dysfunction of“Yang promotes Qi transformation and Yin promotes forming”leads to the coagulation of pathological products such as phlegm and blood stasis in the breast meridian,resulting in the occurrence of acne mastitis.In clinical treatment,we should take“warm Yang to promote Qi transformation”as the treatment principle,and add or subtract according to the specific situation.
