Small peptides function as key signals in processes,such as plant cell differentiation,organ development and defenses to biotic stresses.A large number of small peptide precursor genes have been predicted from the ana...Small peptides function as key signals in processes,such as plant cell differentiation,organ development and defenses to biotic stresses.A large number of small peptide precursor genes have been predicted from the analysis of the soybean(Glycine max) whole genome DNA sequence.However,most of these genes have unknown characteristics and functions.In this report,we systemically searched for the gene families of small peptide precursors that are up-regulated in soybean nitrogen-fixing root nodules.We found 212 genes(encoding peptides shorter than 150 amino acids) that were up-regulated,and among them,79 genes belong to 38 multiple-gene families,but the other 133 genes are unique.Twenty-eight of 38 families are conserved in Arabidopsis,but the other 10 only exist in legumes.We also identified 16 out of the 38 members of the wound-induced polypeptide(WIP) gene family to be upregulated in nitrogen-fixing nodules.We further analyzed homologs of WIP genes in Medicago,Lotus,Arabidopsis and Oryza species and found that a few homologous genes from Medicago truncatula and Lotus japonicus were also upregulated in their nodules and some WIP genes were induced by specific fungal pathogens on soybean and rice.Structure prediction indicated that all WIP prepropeptides contain a conserved DUF3774 domain(including two hydrophobic regions) and most of them have an N-terminal signal sequence.Fluorescence microscopy analysis of two WIP prepropeptides fused to GFP revealed that these proteins are located on the plasma membrane of tobacco leaf cells.Interestingly,34 soybean WIP genes are clustered onto three soybean chromosomes,different from known peptide gene families(such as CLE).Among them,11 highly identical genes are aligned on the 6th chromosome,12 on the 12th,and 11 on the 13th chromosomes.Most of WIP genes from the 12th chromosome share the highest identities with their homologs on the 13th chromosome,suggesting that ancestral WIP genes could have originated from the 13th chromosome,then spread onto the 12th chromosome by chrom展开更多
The radioisotopic tracing method of Hirulog-like peptide（HLP） was established, wihch could be a reference for the pharmacokinetic analysis in small molecule polypeptide research.（）125I-HLP was prepared by Iodogen ...The radioisotopic tracing method of Hirulog-like peptide（HLP） was established, wihch could be a reference for the pharmacokinetic analysis in small molecule polypeptide research.（）125I-HLP was prepared by Iodogen method.The radiochemical purity and stability in vitro of（）125I-HLP were determined by HPLC and SDS-PAGE.The radiochemical purity of(（）125I-HLP) was shown above 95% and was stable for 53 hours.It is the first time using SPE to separate the labeled compound from radioiodide and small molecule polypeptides.The HLP recovery was（92.60±6.31）% by SPE and the coefficient of variation was lower than 10% in a concentration range from 10 to 100 000 ng/mL.The detectable limitation reached 52.79 ng/mL.展开更多
目的观察不同浓度的鱼卵小分子多肽对SD大鼠骨髓间充质干细胞(BM-MSCs)增殖作用的影响。方法分离、培养SD大鼠BM-MSCs,并采用流式细胞仪进行鉴定。选取第3代细胞进行观察和检测。实验组选取25、50、75、100、125、150、200、250、300μg...目的观察不同浓度的鱼卵小分子多肽对SD大鼠骨髓间充质干细胞(BM-MSCs)增殖作用的影响。方法分离、培养SD大鼠BM-MSCs,并采用流式细胞仪进行鉴定。选取第3代细胞进行观察和检测。实验组选取25、50、75、100、125、150、200、250、300μg/ml 9个多肽浓度,对照组加入不含小分子多肽的DMEM完全培养基。各组孵育24 h后采用MTT法检测其细胞增殖情况,确定其最佳作用剂量。在最佳作用量下,于12、16、20、24、32、40、48 h 7个时间点观察鱼卵小分子多肽对SD大鼠BM-MSCs增殖的情况。并采用流式分析鱼卵小分子多肽对BM-MSCs细胞周期的影响。结果鱼卵小分子多肽对BM-MSCs的促增殖作用的最佳浓度为100μg/ml,与对照组相比差异具有统计学意义(P<0.05)。流式分析显示鱼卵小分子多肽可提高SD大鼠BM-MSCs非G1/G0期细胞比例。结论鱼卵小分子多肽对SD大鼠BM-MSCs有促增殖作用,最佳作用浓度为100μg/ml。展开更多
基金supported by National Key Basic Research Program of China(2011CB100702 and 2010CB126501)National Natural Science Foundation of China(31070218)+1 种基金Shanghai Municipal Natural Science Foundation(09ZR1436500)Knowledge Innovation Program of Chinese Academy of Sciences(2009KIP206)
文摘Small peptides function as key signals in processes,such as plant cell differentiation,organ development and defenses to biotic stresses.A large number of small peptide precursor genes have been predicted from the analysis of the soybean(Glycine max) whole genome DNA sequence.However,most of these genes have unknown characteristics and functions.In this report,we systemically searched for the gene families of small peptide precursors that are up-regulated in soybean nitrogen-fixing root nodules.We found 212 genes(encoding peptides shorter than 150 amino acids) that were up-regulated,and among them,79 genes belong to 38 multiple-gene families,but the other 133 genes are unique.Twenty-eight of 38 families are conserved in Arabidopsis,but the other 10 only exist in legumes.We also identified 16 out of the 38 members of the wound-induced polypeptide(WIP) gene family to be upregulated in nitrogen-fixing nodules.We further analyzed homologs of WIP genes in Medicago,Lotus,Arabidopsis and Oryza species and found that a few homologous genes from Medicago truncatula and Lotus japonicus were also upregulated in their nodules and some WIP genes were induced by specific fungal pathogens on soybean and rice.Structure prediction indicated that all WIP prepropeptides contain a conserved DUF3774 domain(including two hydrophobic regions) and most of them have an N-terminal signal sequence.Fluorescence microscopy analysis of two WIP prepropeptides fused to GFP revealed that these proteins are located on the plasma membrane of tobacco leaf cells.Interestingly,34 soybean WIP genes are clustered onto three soybean chromosomes,different from known peptide gene families(such as CLE).Among them,11 highly identical genes are aligned on the 6th chromosome,12 on the 12th,and 11 on the 13th chromosomes.Most of WIP genes from the 12th chromosome share the highest identities with their homologs on the 13th chromosome,suggesting that ancestral WIP genes could have originated from the 13th chromosome,then spread onto the 12th chromosome by chrom
文摘The radioisotopic tracing method of Hirulog-like peptide（HLP） was established, wihch could be a reference for the pharmacokinetic analysis in small molecule polypeptide research.（）125I-HLP was prepared by Iodogen method.The radiochemical purity and stability in vitro of（）125I-HLP were determined by HPLC and SDS-PAGE.The radiochemical purity of(（）125I-HLP) was shown above 95% and was stable for 53 hours.It is the first time using SPE to separate the labeled compound from radioiodide and small molecule polypeptides.The HLP recovery was（92.60±6.31）% by SPE and the coefficient of variation was lower than 10% in a concentration range from 10 to 100 000 ng/mL.The detectable limitation reached 52.79 ng/mL.
文摘目的观察不同浓度的鱼卵小分子多肽对SD大鼠骨髓间充质干细胞(BM-MSCs)增殖作用的影响。方法分离、培养SD大鼠BM-MSCs,并采用流式细胞仪进行鉴定。选取第3代细胞进行观察和检测。实验组选取25、50、75、100、125、150、200、250、300μg/ml 9个多肽浓度,对照组加入不含小分子多肽的DMEM完全培养基。各组孵育24 h后采用MTT法检测其细胞增殖情况,确定其最佳作用剂量。在最佳作用量下,于12、16、20、24、32、40、48 h 7个时间点观察鱼卵小分子多肽对SD大鼠BM-MSCs增殖的情况。并采用流式分析鱼卵小分子多肽对BM-MSCs细胞周期的影响。结果鱼卵小分子多肽对BM-MSCs的促增殖作用的最佳浓度为100μg/ml,与对照组相比差异具有统计学意义(P<0.05)。流式分析显示鱼卵小分子多肽可提高SD大鼠BM-MSCs非G1/G0期细胞比例。结论鱼卵小分子多肽对SD大鼠BM-MSCs有促增殖作用,最佳作用浓度为100μg/ml。
