Curcin, a ribosome-inactivating protein with a molecular weight of about 28.2 kD, which strongly inhibits the protein synthesis in rabbit reticulocyte lysate system with an IC50 value of about (0.19 +/- 0.01) nmol/L, ...Curcin, a ribosome-inactivating protein with a molecular weight of about 28.2 kD, which strongly inhibits the protein synthesis in rabbit reticulocyte lysate system with an IC50 value of about (0.19 +/- 0.01) nmol/L, was purified from the seeds of Jatropha curcas L. The protein has the activity of rRNA N-glycosidase. Degenerate primers were designed based on the N-terminal partial sequence from purified curcin. The full-length curcin cDNA by RT-PCR and 5'-RACE was cloned. The deduced amino acids sequence indicates that a preprotein with 20 amino acid residues is first translated and then processed to a mature protein with 251 amino acids. The deduced amino acids sequence shares homology of 33% and 57% to those of type I ribosome-inactivating proteins (RIPs) and A chain of type II RIPs, respectively. The sequence encoding mature curcin was integrated into the pQE-30 vector for expression in Escherichia coli strain M15 (pREP4). The purified recombinant curcin was able to inhibit protein synthesis in rabbit reticulocyte lysate system.展开更多
Z961215 大麦核糖体失活蛋白基因的克隆及序列分析/吴志浩,朱祯…(中科院遗传所)//农业生物技术学报.-1995,3(4).-64～70以大麦基因组 DNA 为模板,通过合成5′-端及3′-端的一对特异引物,利用多聚酶链式反应(PCR)扩增出了大麦核糖体失...Z961215 大麦核糖体失活蛋白基因的克隆及序列分析/吴志浩,朱祯…(中科院遗传所)//农业生物技术学报.-1995,3(4).-64～70以大麦基因组 DNA 为模板,通过合成5′-端及3′-端的一对特异引物,利用多聚酶链式反应(PCR)扩增出了大麦核糖体失活蛋白编码序列,随后将其克隆到 E.coli 质粒上。序列分析表明,大麦核糖体失活蛋白基因由846个核苷酸组成,编码由281个氨基酸残基组成的多肽。与发表序列相比较,核苷酸序列及推导的氨基酸序列的同源性分别为93.1％和92％。(图5参9)Z961216 西藏裸大麦主要性状的遗传参数研究/王建林,胡书银…(西藏农牧学院农学系)//遗传.-1996,18(2).展开更多
A 1.2kb and 1.3kb DNA fragments were amplified in DNA samples extracted from periwinkle exhibiting yellows naturally. The amplified fragments were cloned, sequenced and compared with other phytoplasma strains. The res...A 1.2kb and 1.3kb DNA fragments were amplified in DNA samples extracted from periwinkle exhibiting yellows naturally. The amplified fragments were cloned, sequenced and compared with other phytoplasma strains. The results showed that the 16S rDNA and ribosomal protein gene of periwinkle yellows phytoplasma consist of 1239bp and 1244bp, respectively. They all shared high homology (higher than 97.5%) with phytoplasma strains of 16S rI group. Thus, the result made it clear that the phytoplasma strain of periwinkle yellows (PY) is one of the members of phytoplasma 16SrI group.展开更多
文摘Curcin, a ribosome-inactivating protein with a molecular weight of about 28.2 kD, which strongly inhibits the protein synthesis in rabbit reticulocyte lysate system with an IC50 value of about (0.19 +/- 0.01) nmol/L, was purified from the seeds of Jatropha curcas L. The protein has the activity of rRNA N-glycosidase. Degenerate primers were designed based on the N-terminal partial sequence from purified curcin. The full-length curcin cDNA by RT-PCR and 5'-RACE was cloned. The deduced amino acids sequence indicates that a preprotein with 20 amino acid residues is first translated and then processed to a mature protein with 251 amino acids. The deduced amino acids sequence shares homology of 33% and 57% to those of type I ribosome-inactivating proteins (RIPs) and A chain of type II RIPs, respectively. The sequence encoding mature curcin was integrated into the pQE-30 vector for expression in Escherichia coli strain M15 (pREP4). The purified recombinant curcin was able to inhibit protein synthesis in rabbit reticulocyte lysate system.
文摘A 1.2kb and 1.3kb DNA fragments were amplified in DNA samples extracted from periwinkle exhibiting yellows naturally. The amplified fragments were cloned, sequenced and compared with other phytoplasma strains. The results showed that the 16S rDNA and ribosomal protein gene of periwinkle yellows phytoplasma consist of 1239bp and 1244bp, respectively. They all shared high homology (higher than 97.5%) with phytoplasma strains of 16S rI group. Thus, the result made it clear that the phytoplasma strain of periwinkle yellows (PY) is one of the members of phytoplasma 16SrI group.
