Arid and semi-arid regions of China account for more than half of the country. Because of drought resistance and high nutritive value, elite foxtail millet (Setaria Italica (L.) P. Beauv.) is one of the most important...Arid and semi-arid regions of China account for more than half of the country. Because of drought resistance and high nutritive value, elite foxtail millet (Setaria Italica (L.) P. Beauv.) is one of the most important cereal crops in China. Evaluation of germplasm and genetic diversity of foxtail millet is still in its infancy, but prolamin could play an important role as a protein marker. To investigate the genetic diversity and population structure of foxtail millet from different ecological zones of China, 90 accessions of foxtail millet were collected from three major ecological areas: North, Northwest, and Northeast China. The prolamin contents were examined by acid polyacrylamide gel electrophoresis (acid-PAGE). Five to twenty-two prolamin bands appeared in tested varieties, of which were polymorphic, so prolamin patterns of foxtail millet varieties can be used in variety identification and evaluation. Structure analysis identified six groups, which matches their pedigree information but not their geographic origins. This indicated a high degree (87.78%) of consistency with a phylogenetic classification based on SSR. The results showed prolamin banding patterns were an effective method for analyzing foxtail millet genetic variability.展开更多
Polamines are the major seed storage proteinsin most of the sereals. The rice prolaminescontain a high mole percentage of glutamineresidues and low levels of lysine, histidine,cystine, and methionine. Among the fourco...Polamines are the major seed storage proteinsin most of the sereals. The rice prolaminescontain a high mole percentage of glutamineresidues and low levels of lysine, histidine,cystine, and methionine. Among the fourcomponent of rice prolamines, 16KDa and10KDa contain more sulfate-riched amino acidresidues. So the total prolamine content andthe percentages of each component were im-portant to parameters for the rice storage pro-tein and its nutrient value. Prolamines of 36rice lines were studied with SDS-PAGE, andrelative quantities of 16 KDa, 13 a, 13 b, and展开更多
Nutritious value of seed storage protein is low due to deficiency in essential amino acid contents. Cereals are mainly deficient in lysine and legumes in sulfur-containing amino acids (methionine and cysteine). So far...Nutritious value of seed storage protein is low due to deficiency in essential amino acid contents. Cereals are mainly deficient in lysine and legumes in sulfur-containing amino acids (methionine and cysteine). So far, several sufur-rich seed protein genes have been isolated and the essential amino acid contents of seed proteins were increased in transgenic tobacco and Brassica napus. In this paper we report the isolation and sequencing of the 10kd prolamin gene from seeds. Poly(A) RNA were prepared from the immature endosperms of japonica rice, Sachiminori, 10 d after flowering. Complementary DNAs were synthesized according to Promega Instruction Manual. Two primers were synthesized and their sequences were Primer Ⅰ: CGTCTACACCATCTGGAATC, Primer Ⅱ: GTGTTTGCAGATAGTATGC. The amplified fragraents were inserted into the Sma I site of pGEM-7zf(+) and was used to transform E. coli JM101 after PCR reaction. DNA sequence were determined by Sanger’s Chain-termination method. Synthesis of cDNA. Using mRNAs as展开更多
The 5' upstream region (—680—+40) containing the complete signal peptide coding sequence of the rice seed storage prolamin gene was amplified in vitro with polymerase chain reaction from the genome of Chinese su...The 5' upstream region (—680—+40) containing the complete signal peptide coding sequence of the rice seed storage prolamin gene was amplified in vitro with polymerase chain reaction from the genome of Chinese super rice cultivar Zhonghua 8. Physical map and DNA sequence analysis showed strong homology with the 5'-flanking region of rice prolamin gene reported by Kim in 1988. No changes in the signal peptide coding sequence and a long leader sequence with several small ORFs were found. Chimeric gene containing 5'-flanking region of the prolamin gene has been transcriptionally fused with the β-glucuronidase reporter gene. The fusion junction was confirmed by both physical map and DNA sequence analysis. The resultant chimeric gene was used to transform the tabacco explants by Ti binary system of Agrobacterium tumefaciens LBA4404. With dot and Southern blotting hybridization, three transgenic tobacco plants with the copies of chimeric GUS genes as many as 20 were obtained. Histochemical analysis revealed that the GUS activity in the endosperm tissues of tobacco seeds at the developmental stage was about 20 DAP. No GUS activity was found in leaves, stems, roots and flowers of the transgenic tobacco plants. Therefore, we concluded that the 5'-upstream cis-elements from—680 to—18 were enough to confer the endosperm-specific and temporal expression of rice prolamin gene.展开更多
Prolamins have attracted increasing attention in recent years because of their potential application in active ingredient encapsulation and food packaging.As a byproduct of the Baijiu brewing industry with huge produc...Prolamins have attracted increasing attention in recent years because of their potential application in active ingredient encapsulation and food packaging.As a byproduct of the Baijiu brewing industry with huge production(40 million tons every year),Jiuzao is a great source of prolamin because the fermentation process consumes mainly starch instead of protein.However,information on the extraction method and the characteristics of prolamin from Baijiu Jiuzao(PBJ)is still lacking at present.In this study,PBJ was extracted and characterized in terms of composition,secondary structure,morphology,aggregation state and solubility.The results suggested that the PBJ consisted of partial fractions of kafirin withoutγ-kafirin.PBJ had moreα-helix structures,was more hydrophobic,and more easily aggregated together than kafirin.Morphology analysis showed that the surface of the PBJ powder consisted of joint spheres or dumbbell particles.Aggregates were formed at the molecular level with Rg and Dmax of 13.94 nm and 52.61 nm,respectively.These findings updated the new knowledge of prolamin from byproducts of the brewing industry and provided an environmentally friendly way to recycle Jiuzao.展开更多
Prolamins(gliadin-related proteins)are cereal proteins which are a source of polypeptides that are toxic to people showing intolerance or allergy to gluten.Therefore,serious safety concerns regarding occasional and ro...Prolamins(gliadin-related proteins)are cereal proteins which are a source of polypeptides that are toxic to people showing intolerance or allergy to gluten.Therefore,serious safety concerns regarding occasional and routine beer consumption emerge due to the presence of immunoreactive amino acid sequences of cereal origin that remain in the final product.The aim of the study was to investigate the presence of prolamins and gluten in samples of beers(72 commercially available beverages)produced in Central Europe.A competitive ELISA and immunoblotting methods were applied to analyze the presence and concentration of immunoreactive proteins/peptides.Gluten level in the studied beers ranged from 10 to over 6000 mg/kg.Approximately 30%beers could be regarded as gluten-free,around 40%were very low gluten and only 25%beers contained considerable amounts of gluten.The highest level of prolamin content was detected in wheat beers.We demonstrated that these high concentrations of prolamins are probably derived from residues of non-degraded wheat gliadins.展开更多
利用A—PAGE（Acid—polyacrylamide gel electrophoresis）技术对来源于32个国家的53份红花材料进行了种子醇溶蛋白检测。结果表明，这些红花材料具有丰富的醇溶蛋白等位变异，共分离出15条迁移率不同的谱带。其中，每份材料可电泳出5...利用A—PAGE（Acid—polyacrylamide gel electrophoresis）技术对来源于32个国家的53份红花材料进行了种子醇溶蛋白检测。结果表明，这些红花材料具有丰富的醇溶蛋白等位变异，共分离出15条迁移率不同的谱带。其中，每份材料可电泳出5-12条谱带，平均8．5条，所有材料有1条共有带。红花种子醇溶蛋白可作为评价红花遗传多样性的工具之一。不同材料的遗传相似系数（GS）变异范围为0．375—1．000，平均值为0．752。聚类分析结果表明，在GS值为0．752的水平上供试材料聚为6大类，其亲缘关系远近与地理来源关系不大。其中，来自中国的7份材料分别被聚在了3个大类中，表明中国红花种子醇溶蛋白遗传多样性比较丰富。展开更多
文摘Arid and semi-arid regions of China account for more than half of the country. Because of drought resistance and high nutritive value, elite foxtail millet (Setaria Italica (L.) P. Beauv.) is one of the most important cereal crops in China. Evaluation of germplasm and genetic diversity of foxtail millet is still in its infancy, but prolamin could play an important role as a protein marker. To investigate the genetic diversity and population structure of foxtail millet from different ecological zones of China, 90 accessions of foxtail millet were collected from three major ecological areas: North, Northwest, and Northeast China. The prolamin contents were examined by acid polyacrylamide gel electrophoresis (acid-PAGE). Five to twenty-two prolamin bands appeared in tested varieties, of which were polymorphic, so prolamin patterns of foxtail millet varieties can be used in variety identification and evaluation. Structure analysis identified six groups, which matches their pedigree information but not their geographic origins. This indicated a high degree (87.78%) of consistency with a phylogenetic classification based on SSR. The results showed prolamin banding patterns were an effective method for analyzing foxtail millet genetic variability.
文摘Polamines are the major seed storage proteinsin most of the sereals. The rice prolaminescontain a high mole percentage of glutamineresidues and low levels of lysine, histidine,cystine, and methionine. Among the fourcomponent of rice prolamines, 16KDa and10KDa contain more sulfate-riched amino acidresidues. So the total prolamine content andthe percentages of each component were im-portant to parameters for the rice storage pro-tein and its nutrient value. Prolamines of 36rice lines were studied with SDS-PAGE, andrelative quantities of 16 KDa, 13 a, 13 b, and
文摘Nutritious value of seed storage protein is low due to deficiency in essential amino acid contents. Cereals are mainly deficient in lysine and legumes in sulfur-containing amino acids (methionine and cysteine). So far, several sufur-rich seed protein genes have been isolated and the essential amino acid contents of seed proteins were increased in transgenic tobacco and Brassica napus. In this paper we report the isolation and sequencing of the 10kd prolamin gene from seeds. Poly(A) RNA were prepared from the immature endosperms of japonica rice, Sachiminori, 10 d after flowering. Complementary DNAs were synthesized according to Promega Instruction Manual. Two primers were synthesized and their sequences were Primer Ⅰ: CGTCTACACCATCTGGAATC, Primer Ⅱ: GTGTTTGCAGATAGTATGC. The amplified fragraents were inserted into the Sma I site of pGEM-7zf(+) and was used to transform E. coli JM101 after PCR reaction. DNA sequence were determined by Sanger’s Chain-termination method. Synthesis of cDNA. Using mRNAs as
基金Project supported by Rockefeller Foundation for International Program on Rice Biotechnology.
文摘The 5' upstream region (—680—+40) containing the complete signal peptide coding sequence of the rice seed storage prolamin gene was amplified in vitro with polymerase chain reaction from the genome of Chinese super rice cultivar Zhonghua 8. Physical map and DNA sequence analysis showed strong homology with the 5'-flanking region of rice prolamin gene reported by Kim in 1988. No changes in the signal peptide coding sequence and a long leader sequence with several small ORFs were found. Chimeric gene containing 5'-flanking region of the prolamin gene has been transcriptionally fused with the β-glucuronidase reporter gene. The fusion junction was confirmed by both physical map and DNA sequence analysis. The resultant chimeric gene was used to transform the tabacco explants by Ti binary system of Agrobacterium tumefaciens LBA4404. With dot and Southern blotting hybridization, three transgenic tobacco plants with the copies of chimeric GUS genes as many as 20 were obtained. Histochemical analysis revealed that the GUS activity in the endosperm tissues of tobacco seeds at the developmental stage was about 20 DAP. No GUS activity was found in leaves, stems, roots and flowers of the transgenic tobacco plants. Therefore, we concluded that the 5'-upstream cis-elements from—680 to—18 were enough to confer the endosperm-specific and temporal expression of rice prolamin gene.
基金supported by the Science and Technology Program of Tibet Autonomous Region,China(XZ202001ZY0017N).
文摘Prolamins have attracted increasing attention in recent years because of their potential application in active ingredient encapsulation and food packaging.As a byproduct of the Baijiu brewing industry with huge production(40 million tons every year),Jiuzao is a great source of prolamin because the fermentation process consumes mainly starch instead of protein.However,information on the extraction method and the characteristics of prolamin from Baijiu Jiuzao(PBJ)is still lacking at present.In this study,PBJ was extracted and characterized in terms of composition,secondary structure,morphology,aggregation state and solubility.The results suggested that the PBJ consisted of partial fractions of kafirin withoutγ-kafirin.PBJ had moreα-helix structures,was more hydrophobic,and more easily aggregated together than kafirin.Morphology analysis showed that the surface of the PBJ powder consisted of joint spheres or dumbbell particles.Aggregates were formed at the molecular level with Rg and Dmax of 13.94 nm and 52.61 nm,respectively.These findings updated the new knowledge of prolamin from byproducts of the brewing industry and provided an environmentally friendly way to recycle Jiuzao.
文摘Prolamins(gliadin-related proteins)are cereal proteins which are a source of polypeptides that are toxic to people showing intolerance or allergy to gluten.Therefore,serious safety concerns regarding occasional and routine beer consumption emerge due to the presence of immunoreactive amino acid sequences of cereal origin that remain in the final product.The aim of the study was to investigate the presence of prolamins and gluten in samples of beers(72 commercially available beverages)produced in Central Europe.A competitive ELISA and immunoblotting methods were applied to analyze the presence and concentration of immunoreactive proteins/peptides.Gluten level in the studied beers ranged from 10 to over 6000 mg/kg.Approximately 30%beers could be regarded as gluten-free,around 40%were very low gluten and only 25%beers contained considerable amounts of gluten.The highest level of prolamin content was detected in wheat beers.We demonstrated that these high concentrations of prolamins are probably derived from residues of non-degraded wheat gliadins.
文摘利用A—PAGE（Acid—polyacrylamide gel electrophoresis）技术对来源于32个国家的53份红花材料进行了种子醇溶蛋白检测。结果表明，这些红花材料具有丰富的醇溶蛋白等位变异，共分离出15条迁移率不同的谱带。其中，每份材料可电泳出5-12条谱带，平均8．5条，所有材料有1条共有带。红花种子醇溶蛋白可作为评价红花遗传多样性的工具之一。不同材料的遗传相似系数（GS）变异范围为0．375—1．000，平均值为0．752。聚类分析结果表明，在GS值为0．752的水平上供试材料聚为6大类，其亲缘关系远近与地理来源关系不大。其中，来自中国的7份材料分别被聚在了3个大类中，表明中国红花种子醇溶蛋白遗传多样性比较丰富。
