High-molecular-weight glutenin subunits (HMW-GSs), one class of seed storage proteins in wheat, play an important role in determining bread-making quality of flour. More and more proves support that HMW-GS- 1Bx 14 a...High-molecular-weight glutenin subunits (HMW-GSs), one class of seed storage proteins in wheat, play an important role in determining bread-making quality of flour. More and more proves support that HMW-GS- 1Bx 14 and - 1Bx 15 subunits are strongly positively associated with good bread-making and excellent noodle-making quality. The two subunits are encoded by two genes, Glu-1Bx14 and Glu-1Bx15, which are tightly linked and located on the 1BL. Protein assay by SDS-PAGE indicated that the expression of Glu-1Bx14 gene was always much stronger than that of Glu-1By15 in the same variety. But, variation of expression level for Glu-1By15 gene existed among varieties, such as in Xiaoyan 54, Xiaoyan 6, Yanzhan 1 and Shanyou 225. We also investigated the transcription difference of Glu-1By14 and Glu-1By15 genes in Xiaoyan 54 and Shanyou 225 by semi-quantitative RT-PCR method. The Glu-1By14 always transcripts much more than the Glu-1By15. This was basically consistent with the translation difference between the two genes. Promoters of 1Bx14, 1By15, 1By8, 1Dx2 and 1Dy12 were cloned from Xiaoyan 54, Chinese Spring and Aegilops tauschii. Sequence analysis indicated that the HMW-GS genes had high homology at their promoter regions. However, significant difference existed between sequence of 1Bx14 promoter and those of other HMW-GS genes. The transient expression experiment showed that the promoter of 1By15 has lower activity than that of 1Bx14, which was consistent with their transcription level of the two genes in varieties. In addition, transient expression of the gus driven by the promoter (P2) of HMW-GS 1Dx2 gene was higher than by other HMW-GS promoters. Therefore, we constructed 1By15 gene expression vector driven by the 1Dx2 promoter, and transformed the 1By15 gene into wheat commercial variety, Jimai 20 by pollen tube method. Of 45 independent transgenic lines identified by PCR, 3 were confirmed to contain the HMW-GS 1By15 gene via Southern hybridization. The delivered 1By15 gene expressed the expected HMW-GS protein in the seeds of transgenic plants.展开更多
The high molecular weight glutenin subunit (HMW-GS) pair 1Bx13+1By16 are recognized to positively correlate with bread-making quality; however, their molecular data remain unknown. In order to reveal the mechanism ...The high molecular weight glutenin subunit (HMW-GS) pair 1Bx13+1By16 are recognized to positively correlate with bread-making quality; however, their molecular data remain unknown. In order to reveal the mechanism by which 1By16 and 1Bx13 creates high quality, their open reading frames (ORFs) were amplified from common wheat Atlas66 and Jimai 20 using primers that were designed based on published sequences of HMW glutenin genes. The ORF of 1By16 was 2220bp, deduced into 738 amino acid residues with seven cysteines including 59 hexapeptides and 22 nanopeptides motifs. The ORF of 1Bx13 was 2385bp, deduced into 795 amino acid residues with four cysteines including 68 hexapeptides, 25 nanopeptides and six tripepUdes motifs. We found that 1By16 was the largest y-type HMW glutenin gene described to date in common wheat. The 1By16 had 36 amino acid residues inserted in the central repetitive domain compared with 1By15. Expression in bacteria and western-blot tests confirmed that the sequence cloned was the ORF of HMW-GS 1By16, and that 1Bx13 was one of the largest 1Bx genes that have been described so far in common wheat, exhibiting a hexapeptide (PGQGQQ) insertion in the end of central repetitive domain compared with 1Bx7. A phylogenetic tree based on the deduced full-length amino acid sequence alignment of the published HMW-GS genes showed that the 1By16 was clustered with Glu-1B-2, and that the 1Bx13 was clustered with Glu-1B-1 alleles.展开更多
背景:类风湿关节炎是以持续性滑膜炎和进行性骨破坏为主要病理特征的慢性自身免疫性疾病。巨噬细胞作为关键的效应细胞,主要通过极化成不同功能表型,在类风湿关节炎发病机制中发挥核心作用。目的:综述巨噬细胞极化在类风湿关节炎骨破坏...背景:类风湿关节炎是以持续性滑膜炎和进行性骨破坏为主要病理特征的慢性自身免疫性疾病。巨噬细胞作为关键的效应细胞,主要通过极化成不同功能表型,在类风湿关节炎发病机制中发挥核心作用。目的:综述巨噬细胞极化在类风湿关节炎骨破坏中的作用及调控机制和最新治疗进展。方法:利用计算机检索Web of Science核心数据库、万方数据库和中国知网2005-2024年期间发表的相关文献。中文检索词为“巨噬细胞,类风湿关节炎,极化,骨与关节,软骨,自身免疫,炎症,M1巨噬细胞,M2巨噬细胞”,英文检索词为“macrophages,rheumatoid arthritis,polarization,bone and Joints,cartilage,autoimmunity,inflammation,M1 macrophage,M2 macrophage”,最终对53篇文献展开综述。结果与结论:类风湿关节炎是一种慢性炎症性疾病,以持续炎症性骨破坏为特征,如果治疗不当,可导致关节畸形,甚至功能丧失。巨噬细胞M1/M2极化比例失衡是类风湿关节炎疾病进展的关键点,这强调了研究巨噬细胞极化在类风湿关节炎炎症性骨破坏中的作用及调控机制的必要性,巨噬细胞极化调控机制能够作为新型治疗剂的潜在靶点。展开更多
文摘High-molecular-weight glutenin subunits (HMW-GSs), one class of seed storage proteins in wheat, play an important role in determining bread-making quality of flour. More and more proves support that HMW-GS- 1Bx 14 and - 1Bx 15 subunits are strongly positively associated with good bread-making and excellent noodle-making quality. The two subunits are encoded by two genes, Glu-1Bx14 and Glu-1Bx15, which are tightly linked and located on the 1BL. Protein assay by SDS-PAGE indicated that the expression of Glu-1Bx14 gene was always much stronger than that of Glu-1By15 in the same variety. But, variation of expression level for Glu-1By15 gene existed among varieties, such as in Xiaoyan 54, Xiaoyan 6, Yanzhan 1 and Shanyou 225. We also investigated the transcription difference of Glu-1By14 and Glu-1By15 genes in Xiaoyan 54 and Shanyou 225 by semi-quantitative RT-PCR method. The Glu-1By14 always transcripts much more than the Glu-1By15. This was basically consistent with the translation difference between the two genes. Promoters of 1Bx14, 1By15, 1By8, 1Dx2 and 1Dy12 were cloned from Xiaoyan 54, Chinese Spring and Aegilops tauschii. Sequence analysis indicated that the HMW-GS genes had high homology at their promoter regions. However, significant difference existed between sequence of 1Bx14 promoter and those of other HMW-GS genes. The transient expression experiment showed that the promoter of 1By15 has lower activity than that of 1Bx14, which was consistent with their transcription level of the two genes in varieties. In addition, transient expression of the gus driven by the promoter (P2) of HMW-GS 1Dx2 gene was higher than by other HMW-GS promoters. Therefore, we constructed 1By15 gene expression vector driven by the 1Dx2 promoter, and transformed the 1By15 gene into wheat commercial variety, Jimai 20 by pollen tube method. Of 45 independent transgenic lines identified by PCR, 3 were confirmed to contain the HMW-GS 1By15 gene via Southern hybridization. The delivered 1By15 gene expressed the expected HMW-GS protein in the seeds of transgenic plants.
基金the National Natural Science Foundation of China (30671293)the High-Tech Research and Development (863) Program of China(2006AA100102).
文摘The high molecular weight glutenin subunit (HMW-GS) pair 1Bx13+1By16 are recognized to positively correlate with bread-making quality; however, their molecular data remain unknown. In order to reveal the mechanism by which 1By16 and 1Bx13 creates high quality, their open reading frames (ORFs) were amplified from common wheat Atlas66 and Jimai 20 using primers that were designed based on published sequences of HMW glutenin genes. The ORF of 1By16 was 2220bp, deduced into 738 amino acid residues with seven cysteines including 59 hexapeptides and 22 nanopeptides motifs. The ORF of 1Bx13 was 2385bp, deduced into 795 amino acid residues with four cysteines including 68 hexapeptides, 25 nanopeptides and six tripepUdes motifs. We found that 1By16 was the largest y-type HMW glutenin gene described to date in common wheat. The 1By16 had 36 amino acid residues inserted in the central repetitive domain compared with 1By15. Expression in bacteria and western-blot tests confirmed that the sequence cloned was the ORF of HMW-GS 1By16, and that 1Bx13 was one of the largest 1Bx genes that have been described so far in common wheat, exhibiting a hexapeptide (PGQGQQ) insertion in the end of central repetitive domain compared with 1Bx7. A phylogenetic tree based on the deduced full-length amino acid sequence alignment of the published HMW-GS genes showed that the 1By16 was clustered with Glu-1B-2, and that the 1Bx13 was clustered with Glu-1B-1 alleles.
文摘背景:类风湿关节炎是以持续性滑膜炎和进行性骨破坏为主要病理特征的慢性自身免疫性疾病。巨噬细胞作为关键的效应细胞,主要通过极化成不同功能表型,在类风湿关节炎发病机制中发挥核心作用。目的:综述巨噬细胞极化在类风湿关节炎骨破坏中的作用及调控机制和最新治疗进展。方法:利用计算机检索Web of Science核心数据库、万方数据库和中国知网2005-2024年期间发表的相关文献。中文检索词为“巨噬细胞,类风湿关节炎,极化,骨与关节,软骨,自身免疫,炎症,M1巨噬细胞,M2巨噬细胞”,英文检索词为“macrophages,rheumatoid arthritis,polarization,bone and Joints,cartilage,autoimmunity,inflammation,M1 macrophage,M2 macrophage”,最终对53篇文献展开综述。结果与结论:类风湿关节炎是一种慢性炎症性疾病,以持续炎症性骨破坏为特征,如果治疗不当,可导致关节畸形,甚至功能丧失。巨噬细胞M1/M2极化比例失衡是类风湿关节炎疾病进展的关键点,这强调了研究巨噬细胞极化在类风湿关节炎炎症性骨破坏中的作用及调控机制的必要性,巨噬细胞极化调控机制能够作为新型治疗剂的潜在靶点。