Mycoplasma genitalium is the main causative agent for non-gonococcal and non-chlamydial urethritis. P32 is the putative surface-exposed membrane protein of M. genitalium and it has substaintial identity in amino acid ...Mycoplasma genitalium is the main causative agent for non-gonococcal and non-chlamydial urethritis. P32 is the putative surface-exposed membrane protein of M. genitalium and it has substaintial identity in amino acid sequence with adhesin protein P30 from M. pneumoniae. Since M. pneumoniae mutants lacking P30 protein is defective in cytadherence, P32 protein has been proposed to be an essential adhesin implicated in the adherence of M. genitaliurn to host cells. The prokaryotic expression vector pET-30 （ ＋ ）/p32 was constructed in the present study, and the recombinant protein was expressed in E. coli and purified under denaturing condition. As demonstrated by the immuno- blotting analysis, the recombinant protein could react with rabbit antisera against M. genitalium, and adherence inhibition assays were performed with antisera against this recombinant protein. It was demonstrated that P32 protein apperared to be an adhesion protein of M. genitalium, thus providing the experimental basis for better understanding of the pathogenesis of M. genitalium infection and for the development of the related vaccines against the infection.展开更多
The opioid receptor-libel receptor (ORL), an orphan receptor whose human and murine complementary DNAs,has been characterized recently. ORL transcripts are particularly abundant in the central nervous system. We demon...The opioid receptor-libel receptor (ORL), an orphan receptor whose human and murine complementary DNAs,has been characterized recently. ORL transcripts are particularly abundant in the central nervous system. We demonstrated that ORL expressed in human neuroblastoma SK-N-SH and SH-SY5Y cell lines by radioligand binding assay, reverse transcription polymerase chain reaction (RT-PCR) and Northern analysis in the present study. Stimulation with ORL1 specific agonist, nociceptin/orphanin Fo, increased [34S]GTPrγS binding to SK-N-SH cell membranes (EC50 = 14 ±0.45 nM), and attenuated forskolin-stimulated accumulation of cellular cAMP (EC50= 0.80 ±0.45 nM, indicative that activation of ORL1 activates G proteins and inhibits adenylyl cyclase. Activation of ORL1 receptor was also accessed using CHO:hORL1 cell line by microphysiometer. Treatment of nociceptin/orphanin FQ increased extracellular acidification rate significantly.展开更多
The expression and function in growth and apoptosis of the renin-angiotensin system(RAS)was evaluated inhuman glioblastoma.Renin and angiotensinogen(AGT)mRNAs and proteins were found by in situ hybridisationand immuno...The expression and function in growth and apoptosis of the renin-angiotensin system(RAS)was evaluated inhuman glioblastoma.Renin and angiotensinogen(AGT)mRNAs and proteins were found by in situ hybridisationand immunohistochemistry in glioblastoma cells.Angiotensinogen was present in glioblastoma cystic fluids.Thus,human glioblastoma cells produce renin and AGT and secrete AGT.Human glioblastoma and glioblastoma cellsexpressed renin,AGT,renin receptor,AT(2)and/or AT(1)mRNAs and proteins determined by RT-PCR and/展开更多
Expression of opioid receptor-like receptor (ORL1)and its endogenous peptide agonist nociceptin/orphaninFo (N/OFQ) during mouse embryogenesis have been investigated. Transcripts of ORL1 and N/OFQ were detected by RT-P...Expression of opioid receptor-like receptor (ORL1)and its endogenous peptide agonist nociceptin/orphaninFo (N/OFQ) during mouse embryogenesis have been investigated. Transcripts of ORL1 and N/OFQ were detected by RT-PCR in mouse brain of day 8 embryo (E8)and the expression continued afterwards. Northern blotanalysis revealed abundant expression of ORL1 at postnatal day 1 (P1) and N/OFQ at E17 and P1 in the brain butnone was detected in other embryonic tissues. The presence of functional ORL1 in mouse embryonic brain wasalso confirmed by specific binding of [3H] N/OFQ (kd=1.3±0.5 nM and Bmax = 72±9 fmol/mg protein) as wellas by N/OFQ-stimulated G protein activation.展开更多
Fiber cell initiation is a complex process involving many pathways,including phytohormones and components for transcriptional and posttranscriptional regulation.Here we report expression
Based on the Prototype Theory,the prototypical feature of advertisement is found to be the combination of three language functions:the informative function,the expressive function,and the vocative function.The adverti...Based on the Prototype Theory,the prototypical feature of advertisement is found to be the combination of three language functions:the informative function,the expressive function,and the vocative function.The advertisement translation means the adjustment of the informative function and the expressive function according to the differences between languages or cultures in order to maximize the vocative function.The faithful translation is the closest to the prototype of the source text but not necessarily the best translation.展开更多
To establish a rapid quantification method for heparinase I during its production in recombinant Escherichia coli, a translational fusion vector was constructed by fusing the N terminus of heparinase I to the C termin...To establish a rapid quantification method for heparinase I during its production in recombinant Escherichia coli, a translational fusion vector was constructed by fusing the N terminus of heparinase I to the C terminus of a green fluorescent protein mutant （GFPmutl）. As a result, not only was the functional recombinant expression of heparinase I in E. coli accomplished, but also a linear correlation was obtained between the GFP fluorescence intensity and heparinase I activity, allowing enzyme activity to be quantified rapidly during the fermentation.展开更多
为了研究蒙古马常规冷冻精液生产前后精子中蛋白质的表达变化,试验采用数据非依赖采集(data independent acquisition,DIA)定量蛋白质组学技术对3匹蒙古马精液常规冷冻前后(新鲜精液和冷冻精液)的总蛋白进行定量分析,并将总蛋白在碳水...为了研究蒙古马常规冷冻精液生产前后精子中蛋白质的表达变化,试验采用数据非依赖采集(data independent acquisition,DIA)定量蛋白质组学技术对3匹蒙古马精液常规冷冻前后(新鲜精液和冷冻精液)的总蛋白进行定量分析,并将总蛋白在碳水化合物酶(carbohydrate-active enzyme,CAZyme)、京都基因与基因百科全书同源性(Kyoto encyclopedia of proteins and genomes ortholog,KO)、基因本体论(gene ontology,GO)、真核生物同源类群(eukaryotic orthologous groups,KOG)数据库中注释;以lb(冷冻精液总蛋白的数量/新鲜精液总蛋白的数量)>2或<0.5、P<0.05筛选差异表达蛋白质(differentially expressed proteins,DEPs),对筛选到的DEPs进行GO富集分析和KEGG信号通路分析,最后对部分DEPs进行Western-blot验证。结果表明:共得到新鲜精液和冻精总蛋白数量为2429个,这些蛋白在CAZyme、KO、GO、KOG数据库分别注释到64,923,2028,1868个,分别占总蛋白数量的2.63%、38.00%、83.49%和76.90%;共筛选出1406个DEPs,其中下调DEPs为794个,上调DEPs为612个。在GO功能富集分析中,有369个下调DEPs参与了生物学进程中的生物调控,419个下调DEPs定位于细胞组分中的细胞外区域,631个下调DEPs参与到分子功能中分子功能中,31个上调DEPs参与了生物学进程中的精子发生,161个下调DEPs定位于细胞组分中的线粒体中。在KEGG通路富集分析中,有288个DEPs显著富集代谢通路中。DEPs中膜联蛋白(annexin,ANXA2)、微管蛋白α链(tubulin alpha chain,TUBA8)、精子尾部外致密纤维2(outer dense fiber of sperm tails 2,ODF2)和精子发生相关48(spermatogenesis associated 48,SPATA48)、热休克蛋白70(heat shock protein 70,HSP70)和热休克蛋白90α家族B类成员1(heat shock protein 90-alpha family class B member 1,HSP90AB1)等蛋白与精子密切相关;经Western-blot检测,HSP70和HSP90蛋白在冻精中的表达量极显著低于新鲜精液(P<0.01)。说明冻融过程严重影响了蒙古马精子中HSP70和HSP90蛋白的表达。展开更多
基金National Natural Science Foundation of China(No.30570093).
文摘Mycoplasma genitalium is the main causative agent for non-gonococcal and non-chlamydial urethritis. P32 is the putative surface-exposed membrane protein of M. genitalium and it has substaintial identity in amino acid sequence with adhesin protein P30 from M. pneumoniae. Since M. pneumoniae mutants lacking P30 protein is defective in cytadherence, P32 protein has been proposed to be an essential adhesin implicated in the adherence of M. genitaliurn to host cells. The prokaryotic expression vector pET-30 （ ＋ ）/p32 was constructed in the present study, and the recombinant protein was expressed in E. coli and purified under denaturing condition. As demonstrated by the immuno- blotting analysis, the recombinant protein could react with rabbit antisera against M. genitalium, and adherence inhibition assays were performed with antisera against this recombinant protein. It was demonstrated that P32 protein apperared to be an adhesion protein of M. genitalium, thus providing the experimental basis for better understanding of the pathogenesis of M. genitalium infection and for the development of the related vaccines against the infection.
文摘The opioid receptor-libel receptor (ORL), an orphan receptor whose human and murine complementary DNAs,has been characterized recently. ORL transcripts are particularly abundant in the central nervous system. We demonstrated that ORL expressed in human neuroblastoma SK-N-SH and SH-SY5Y cell lines by radioligand binding assay, reverse transcription polymerase chain reaction (RT-PCR) and Northern analysis in the present study. Stimulation with ORL1 specific agonist, nociceptin/orphanin Fo, increased [34S]GTPrγS binding to SK-N-SH cell membranes (EC50 = 14 ±0.45 nM), and attenuated forskolin-stimulated accumulation of cellular cAMP (EC50= 0.80 ±0.45 nM, indicative that activation of ORL1 activates G proteins and inhibits adenylyl cyclase. Activation of ORL1 receptor was also accessed using CHO:hORL1 cell line by microphysiometer. Treatment of nociceptin/orphanin FQ increased extracellular acidification rate significantly.
文摘The expression and function in growth and apoptosis of the renin-angiotensin system(RAS)was evaluated inhuman glioblastoma.Renin and angiotensinogen(AGT)mRNAs and proteins were found by in situ hybridisationand immunohistochemistry in glioblastoma cells.Angiotensinogen was present in glioblastoma cystic fluids.Thus,human glioblastoma cells produce renin and AGT and secrete AGT.Human glioblastoma and glioblastoma cellsexpressed renin,AGT,renin receptor,AT(2)and/or AT(1)mRNAs and proteins determined by RT-PCR and/
文摘Expression of opioid receptor-like receptor (ORL1)and its endogenous peptide agonist nociceptin/orphaninFo (N/OFQ) during mouse embryogenesis have been investigated. Transcripts of ORL1 and N/OFQ were detected by RT-PCR in mouse brain of day 8 embryo (E8)and the expression continued afterwards. Northern blotanalysis revealed abundant expression of ORL1 at postnatal day 1 (P1) and N/OFQ at E17 and P1 in the brain butnone was detected in other embryonic tissues. The presence of functional ORL1 in mouse embryonic brain wasalso confirmed by specific binding of [3H] N/OFQ (kd=1.3±0.5 nM and Bmax = 72±9 fmol/mg protein) as wellas by N/OFQ-stimulated G protein activation.
文摘Fiber cell initiation is a complex process involving many pathways,including phytohormones and components for transcriptional and posttranscriptional regulation.Here we report expression
文摘Based on the Prototype Theory,the prototypical feature of advertisement is found to be the combination of three language functions:the informative function,the expressive function,and the vocative function.The advertisement translation means the adjustment of the informative function and the expressive function according to the differences between languages or cultures in order to maximize the vocative function.The faithful translation is the closest to the prototype of the source text but not necessarily the best translation.
基金Supported by the National Natural Science Foundation of China (No.20336010 and No.20176025).
文摘To establish a rapid quantification method for heparinase I during its production in recombinant Escherichia coli, a translational fusion vector was constructed by fusing the N terminus of heparinase I to the C terminus of a green fluorescent protein mutant （GFPmutl）. As a result, not only was the functional recombinant expression of heparinase I in E. coli accomplished, but also a linear correlation was obtained between the GFP fluorescence intensity and heparinase I activity, allowing enzyme activity to be quantified rapidly during the fermentation.
文摘为了研究蒙古马常规冷冻精液生产前后精子中蛋白质的表达变化,试验采用数据非依赖采集(data independent acquisition,DIA)定量蛋白质组学技术对3匹蒙古马精液常规冷冻前后(新鲜精液和冷冻精液)的总蛋白进行定量分析,并将总蛋白在碳水化合物酶(carbohydrate-active enzyme,CAZyme)、京都基因与基因百科全书同源性(Kyoto encyclopedia of proteins and genomes ortholog,KO)、基因本体论(gene ontology,GO)、真核生物同源类群(eukaryotic orthologous groups,KOG)数据库中注释;以lb(冷冻精液总蛋白的数量/新鲜精液总蛋白的数量)>2或<0.5、P<0.05筛选差异表达蛋白质(differentially expressed proteins,DEPs),对筛选到的DEPs进行GO富集分析和KEGG信号通路分析,最后对部分DEPs进行Western-blot验证。结果表明:共得到新鲜精液和冻精总蛋白数量为2429个,这些蛋白在CAZyme、KO、GO、KOG数据库分别注释到64,923,2028,1868个,分别占总蛋白数量的2.63%、38.00%、83.49%和76.90%;共筛选出1406个DEPs,其中下调DEPs为794个,上调DEPs为612个。在GO功能富集分析中,有369个下调DEPs参与了生物学进程中的生物调控,419个下调DEPs定位于细胞组分中的细胞外区域,631个下调DEPs参与到分子功能中分子功能中,31个上调DEPs参与了生物学进程中的精子发生,161个下调DEPs定位于细胞组分中的线粒体中。在KEGG通路富集分析中,有288个DEPs显著富集代谢通路中。DEPs中膜联蛋白(annexin,ANXA2)、微管蛋白α链(tubulin alpha chain,TUBA8)、精子尾部外致密纤维2(outer dense fiber of sperm tails 2,ODF2)和精子发生相关48(spermatogenesis associated 48,SPATA48)、热休克蛋白70(heat shock protein 70,HSP70)和热休克蛋白90α家族B类成员1(heat shock protein 90-alpha family class B member 1,HSP90AB1)等蛋白与精子密切相关;经Western-blot检测,HSP70和HSP90蛋白在冻精中的表达量极显著低于新鲜精液(P<0.01)。说明冻融过程严重影响了蒙古马精子中HSP70和HSP90蛋白的表达。
