Nicotinic acetylcholine receptors (nAChRs) play a significant role in excitatory synaptic transmission in insects and are the target for chloronicotinyl and nereistoxin insecticides.In recent years,Chilo suppressalis,...Nicotinic acetylcholine receptors (nAChRs) play a significant role in excitatory synaptic transmission in insects and are the target for chloronicotinyl and nereistoxin insecticides.In recent years,Chilo suppressalis,an economically important pest of rice,developed high resistance against monosultap,a nereistoxin insecticide acting on nAChR.In order to reveal the hypothesized target insensitive mechanism,studies on the molecular property of nAChR from Chilo suppressalis are required.In this study,the full length cDNA of nAChR α subunit from this pest was cloned by RT-PCR.Sequence analysis shows that it is a novel nAChR α subunit,which was named as Cs α 1(Genbank accession No.AF418987).It contains 1?997?bp nucleotides and involves an open reading frame (ORF) encoding a mature protein of 509 amino acids excluding a signal peptide of 24 amino acids.The deduced amino acid sequence was 52%-94% identical to the reported insect nAChR genes.展开更多
[Objective]The α subunit gene of phycobiliprotein from Spirulina maxima was studied in order to provide a basis for the subsequent study of phycobiliprotein.[Method] Amino acids composition,signal peptides,hydrophobi...[Objective]The α subunit gene of phycobiliprotein from Spirulina maxima was studied in order to provide a basis for the subsequent study of phycobiliprotein.[Method] Amino acids composition,signal peptides,hydrophobicity/hydrophilicity and trails-membrane topological structure of α subunit gene of phycobiliprotein from Spirulina maxima which registered in GenBank（GenBank AF441177） were analyzed and predicted by the tools of bioinformatic analysis.Meanwhile,phylogenetic tree was constructed based on α subunit gene of phycobiliprotein from Spirulina maxima,and its molecular evolution was also analyzed.[Result]The phycobiliprotein was rich in amino acids,which not only contained 18 kinds of essential amino acids,but also contained some non-essential amino acids like glycine,aspartic acid,etc.;Analysis on signal peptides and trails-membrane topological structure showed that the phycobiliprotein belonged to intracellular protein;Analysis on hydrophobicity/hydrophilicity showed that the phycobiliprotein belonged to hydrophilic protein;Phylogenetic analysis showed that the phycobiliprotein had a high homology with Arthrospira,which reached 99%-100%.[Conclusion]The study provided a certain reference for studying the relationship and interaction between α subunit and β subunit.展开更多
Heterotrimeric G proteins are known to function as messengers in numerous signal transduction pathways.The nullmutation of RGA(rice heterotrimeric G protein α subunit),which encodes the α subunit of heterotrimeric G...Heterotrimeric G proteins are known to function as messengers in numerous signal transduction pathways.The nullmutation of RGA(rice heterotrimeric G protein α subunit),which encodes the α subunit of heterotrimeric G proteinin rice,causes severe dwarfism and reduced responsiveness to gibberellic acid in rice.However,less is known aboutheterotrimeric G protein in brassinosteroid(BR)signaling,one of the well-understood phytohormone pathways.In thepresent study,we used root elongation inhibition assay,lamina inclination assay and coleoptile elongation analysis todemonstrated reduced sensitivity of dl mutant plants(caused by the null mutation of RGA)to 24-epibrassinolide(24-epiBL),which belongs to brassinosteroids and plays a wide variety of roles in plant growth and development.Moreover,RGA transcript level was decreased in 24-epiBL-treated seedlings in a dose-dependent manner.Our results show thatRGA is involved in rice brassinosteroid response,which may be beneficial to elucidate the molecular mechanisms of Gprotein signaling and provide a novel perspective to understand BR signaling in higher plants.展开更多
The cDNA encoding the G protein αq subunit was isolated from the antennae of Helicoverpa assulta （Guen6e） by reverse transcription polymerase chain reaction （RT-PCR） and named as HassGαq. Sequencing analysis sho...The cDNA encoding the G protein αq subunit was isolated from the antennae of Helicoverpa assulta （Guen6e） by reverse transcription polymerase chain reaction （RT-PCR） and named as HassGαq. Sequencing analysis showed that the fulllength of HassGαq open reading frame （ORF） is 1 062 bp, 353 amino acid residues are encoded. The predicted molecular weights （MW） and isoelectric point （PI） are 41.5 kD and 5.15, respectively. HassGαq gene was then constructed into expression vector pGEX-4T-2 for over expression in prokaryotic cells. The SDS-PAGE and Western blot analysis showed that induced by Isopropyl-β-D-Thiogalactoside （IPTG）, the GST-HassGαq fusion protein is expressed in Escherichia coil BL21, and its MW was found to be about 66 kD nearly equal to the predicted. In addition, RT-PCR analysis showed that the expressions of HassGαq are not tissue specific.展开更多
By PCR method, apo phycoerythrocyanin α subunit gene (pecA) of Mastigocladus laminosus (M. laminosus) was amplified from its genomic DNA, and then cloned in pBluescript. The pecA gene was subcloned into the exp...By PCR method, apo phycoerythrocyanin α subunit gene (pecA) of Mastigocladus laminosus (M. laminosus) was amplified from its genomic DNA, and then cloned in pBluescript. The pecA gene was subcloned into the expression vector pGEMD, and then transformed into E.coli BL21 (DE3). After induction, a new protein of molecular weight 19×10 3 existing in inclusion body was overexpressed. The expressed product was confirmed to be apo phycoerythrocyanin α subunit by Dot ELISA.展开更多
GABAergic input to Gonadotropin-releasing hormone (GnRH) neurons is necessary to initiate the onset of puberty and its action mainly depends on GABAA receptor of which the subunit composition, properties and consequen...GABAergic input to Gonadotropin-releasing hormone (GnRH) neurons is necessary to initiate the onset of puberty and its action mainly depends on GABAA receptor of which the subunit composition, properties and consequently function varies during this period. Nourishing “Yin”-Removing “Fire” Chinese herb mixture, a Chinese herb-based formulation, has been proved that it may retard the initiation of pubertal development in female precocious puberty rats. Our objective is to investigate the effects of Nourishing “Yin”-Removing “Fire” Chinese herb mixture on the expression of GABAA receptor α subunits in hypothalamus. Female Sprague-Dawley rats were divided into normal (N), precocious puberty model (M) induced by danazol, model exposed to saline (MS) and model exposed to Chinese herb mixture (CHM) groups. All rats were administered by the Chinese herb mixture from P15 on. Coefficients of reproductive organs and serum gonadotropins and estradiol levels in M were significantly enhanced while they were significantly decreased in CHM. The hypothalamic GnRH mRNA was also significantly increased in M and in CHM, as well as ERα mRNA. At the mean time, the hypothalamic GABAA receptor α1 and α3 subunits mRNA were more significantly decreased in M than those of N, while they were more significantly enhanced in CHM than those in M (p 0.01), the protein expression of which in hypothalamus had the same trend as the mRNA expression. The evidence suggests that Nourishing “Yin”-Removing “Fire” Chinese herb mixture could significantly retard the sexual development of the precocious rats, and up-regulate the expressions of hypothalamic GABAA receptor α1 and α3 subunits. Our result indicated that GABAA receptor α1 and α3 subunits might involve in the effective treatment of herb mixture on idiopathic precocious puberty.展开更多
[Objective] The paper was to develop genetic engineering vaccine that can express α exotoxin antigen protein efficiently without destroying its immunogenicity for preventing and controlling the diseases caused by Clo...[Objective] The paper was to develop genetic engineering vaccine that can express α exotoxin antigen protein efficiently without destroying its immunogenicity for preventing and controlling the diseases caused by Clostridium perfringens. [Method] Efficiently expressed soluble recombinant α protein was obtained from Escherichia coli expression system by optimizing codon,removing signal peptide,selecting sequences with better hydrophilicity and antigenicity,and optimizing expression conditions. [Result] Mice obtained higher serum antibody level when immunized by α protein,and the immune protection rates against type A,type B,type C and type D C. perfringens were 100%,90%,85% and 90%,respectively. The antibody titer of mice within 7-14 d after the third immunization reached the peak. [Conclusion]The α protein has good immunogenicity,and can be further used to develop genetic engineering subunit vaccines for preventing C. perfringens.展开更多
结合模糊识别方法评价的数学模型法和联合国粮农组织/世界卫生组织（Food and Agriculture Organiz ation/World Health Organization,FAO/WHO）、鸡蛋蛋白两种模式下的化学分析法评价两个致敏蛋白α亚基缺失型大豆蛋白质氨基酸营养价值...结合模糊识别方法评价的数学模型法和联合国粮农组织/世界卫生组织（Food and Agriculture Organiz ation/World Health Organization,FAO/WHO）、鸡蛋蛋白两种模式下的化学分析法评价两个致敏蛋白α亚基缺失型大豆蛋白质氨基酸营养价值,解析α亚基缺失特性对大豆氨基酸组分及营养品质的影响。用氨基酸分析仪测定氨基酸的组分和含量,α亚基缺失特性用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecylsulfate polyacrylamide gel electrophoresis,SDS-PAGE）确认,蛋白和脂肪含量用Perten 8620近红外谷物分析仪测定。结果表明：1）致敏蛋白α亚基缺失型大豆的氨基酸总量、必需氨基酸总量、蛋白质、油分含量不因α亚基的缺失而降低;2）致敏蛋白α亚基缺失型大豆的11S/7S比值在4.65以上,高于目前普遍报道的2.0~3.0;3）致敏蛋白α亚基缺失型大豆必需氨基酸含量接近或高于FAO/WHO标准;两种模式下,α亚基缺失型大豆的5种化学评分及贴进度值都很高,接近标准蛋白。致敏蛋白α亚基缺失型大豆蛋白质氨基酸组分平衡,11S/7S比值更优,营养品质更高。展开更多
文摘Nicotinic acetylcholine receptors (nAChRs) play a significant role in excitatory synaptic transmission in insects and are the target for chloronicotinyl and nereistoxin insecticides.In recent years,Chilo suppressalis,an economically important pest of rice,developed high resistance against monosultap,a nereistoxin insecticide acting on nAChR.In order to reveal the hypothesized target insensitive mechanism,studies on the molecular property of nAChR from Chilo suppressalis are required.In this study,the full length cDNA of nAChR α subunit from this pest was cloned by RT-PCR.Sequence analysis shows that it is a novel nAChR α subunit,which was named as Cs α 1(Genbank accession No.AF418987).It contains 1?997?bp nucleotides and involves an open reading frame (ORF) encoding a mature protein of 509 amino acids excluding a signal peptide of 24 amino acids.The deduced amino acid sequence was 52%-94% identical to the reported insect nAChR genes.
基金Supported by the Special Project for Public-welfare Industry of Chinese Forestry (200704025) The Key Project of Chinese Ministry of Education (102023)~~
文摘[Objective]The α subunit gene of phycobiliprotein from Spirulina maxima was studied in order to provide a basis for the subsequent study of phycobiliprotein.[Method] Amino acids composition,signal peptides,hydrophobicity/hydrophilicity and trails-membrane topological structure of α subunit gene of phycobiliprotein from Spirulina maxima which registered in GenBank（GenBank AF441177） were analyzed and predicted by the tools of bioinformatic analysis.Meanwhile,phylogenetic tree was constructed based on α subunit gene of phycobiliprotein from Spirulina maxima,and its molecular evolution was also analyzed.[Result]The phycobiliprotein was rich in amino acids,which not only contained 18 kinds of essential amino acids,but also contained some non-essential amino acids like glycine,aspartic acid,etc.;Analysis on signal peptides and trails-membrane topological structure showed that the phycobiliprotein belonged to intracellular protein;Analysis on hydrophobicity/hydrophilicity showed that the phycobiliprotein belonged to hydrophilic protein;Phylogenetic analysis showed that the phycobiliprotein had a high homology with Arthrospira,which reached 99%-100%.[Conclusion]The study provided a certain reference for studying the relationship and interaction between α subunit and β subunit.
基金This project was supported by the Major State Basic Research Program of China （2005CB 120806）, National Natural Science Foundation of China for Distinguished Young Scholars （30525026） and the State Transgenic Plant Project （JY04-A-01）
文摘Heterotrimeric G proteins are known to function as messengers in numerous signal transduction pathways.The nullmutation of RGA(rice heterotrimeric G protein α subunit),which encodes the α subunit of heterotrimeric G proteinin rice,causes severe dwarfism and reduced responsiveness to gibberellic acid in rice.However,less is known aboutheterotrimeric G protein in brassinosteroid(BR)signaling,one of the well-understood phytohormone pathways.In thepresent study,we used root elongation inhibition assay,lamina inclination assay and coleoptile elongation analysis todemonstrated reduced sensitivity of dl mutant plants(caused by the null mutation of RGA)to 24-epibrassinolide(24-epiBL),which belongs to brassinosteroids and plays a wide variety of roles in plant growth and development.Moreover,RGA transcript level was decreased in 24-epiBL-treated seedlings in a dose-dependent manner.Our results show thatRGA is involved in rice brassinosteroid response,which may be beneficial to elucidate the molecular mechanisms of Gprotein signaling and provide a novel perspective to understand BR signaling in higher plants.
文摘The cDNA encoding the G protein αq subunit was isolated from the antennae of Helicoverpa assulta （Guen6e） by reverse transcription polymerase chain reaction （RT-PCR） and named as HassGαq. Sequencing analysis showed that the fulllength of HassGαq open reading frame （ORF） is 1 062 bp, 353 amino acid residues are encoded. The predicted molecular weights （MW） and isoelectric point （PI） are 41.5 kD and 5.15, respectively. HassGαq gene was then constructed into expression vector pGEX-4T-2 for over expression in prokaryotic cells. The SDS-PAGE and Western blot analysis showed that induced by Isopropyl-β-D-Thiogalactoside （IPTG）, the GST-HassGαq fusion protein is expressed in Escherichia coil BL21, and its MW was found to be about 66 kD nearly equal to the predicted. In addition, RT-PCR analysis showed that the expressions of HassGαq are not tissue specific.
文摘By PCR method, apo phycoerythrocyanin α subunit gene (pecA) of Mastigocladus laminosus (M. laminosus) was amplified from its genomic DNA, and then cloned in pBluescript. The pecA gene was subcloned into the expression vector pGEMD, and then transformed into E.coli BL21 (DE3). After induction, a new protein of molecular weight 19×10 3 existing in inclusion body was overexpressed. The expressed product was confirmed to be apo phycoerythrocyanin α subunit by Dot ELISA.
文摘GABAergic input to Gonadotropin-releasing hormone (GnRH) neurons is necessary to initiate the onset of puberty and its action mainly depends on GABAA receptor of which the subunit composition, properties and consequently function varies during this period. Nourishing “Yin”-Removing “Fire” Chinese herb mixture, a Chinese herb-based formulation, has been proved that it may retard the initiation of pubertal development in female precocious puberty rats. Our objective is to investigate the effects of Nourishing “Yin”-Removing “Fire” Chinese herb mixture on the expression of GABAA receptor α subunits in hypothalamus. Female Sprague-Dawley rats were divided into normal (N), precocious puberty model (M) induced by danazol, model exposed to saline (MS) and model exposed to Chinese herb mixture (CHM) groups. All rats were administered by the Chinese herb mixture from P15 on. Coefficients of reproductive organs and serum gonadotropins and estradiol levels in M were significantly enhanced while they were significantly decreased in CHM. The hypothalamic GnRH mRNA was also significantly increased in M and in CHM, as well as ERα mRNA. At the mean time, the hypothalamic GABAA receptor α1 and α3 subunits mRNA were more significantly decreased in M than those of N, while they were more significantly enhanced in CHM than those in M (p 0.01), the protein expression of which in hypothalamus had the same trend as the mRNA expression. The evidence suggests that Nourishing “Yin”-Removing “Fire” Chinese herb mixture could significantly retard the sexual development of the precocious rats, and up-regulate the expressions of hypothalamic GABAA receptor α1 and α3 subunits. Our result indicated that GABAA receptor α1 and α3 subunits might involve in the effective treatment of herb mixture on idiopathic precocious puberty.
基金Supported by the 13th Five-Year National Key Research and Development Program(2016YFD0500901)
文摘[Objective] The paper was to develop genetic engineering vaccine that can express α exotoxin antigen protein efficiently without destroying its immunogenicity for preventing and controlling the diseases caused by Clostridium perfringens. [Method] Efficiently expressed soluble recombinant α protein was obtained from Escherichia coli expression system by optimizing codon,removing signal peptide,selecting sequences with better hydrophilicity and antigenicity,and optimizing expression conditions. [Result] Mice obtained higher serum antibody level when immunized by α protein,and the immune protection rates against type A,type B,type C and type D C. perfringens were 100%,90%,85% and 90%,respectively. The antibody titer of mice within 7-14 d after the third immunization reached the peak. [Conclusion]The α protein has good immunogenicity,and can be further used to develop genetic engineering subunit vaccines for preventing C. perfringens.
文摘结合模糊识别方法评价的数学模型法和联合国粮农组织/世界卫生组织（Food and Agriculture Organiz ation/World Health Organization,FAO/WHO）、鸡蛋蛋白两种模式下的化学分析法评价两个致敏蛋白α亚基缺失型大豆蛋白质氨基酸营养价值,解析α亚基缺失特性对大豆氨基酸组分及营养品质的影响。用氨基酸分析仪测定氨基酸的组分和含量,α亚基缺失特性用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecylsulfate polyacrylamide gel electrophoresis,SDS-PAGE）确认,蛋白和脂肪含量用Perten 8620近红外谷物分析仪测定。结果表明：1）致敏蛋白α亚基缺失型大豆的氨基酸总量、必需氨基酸总量、蛋白质、油分含量不因α亚基的缺失而降低;2）致敏蛋白α亚基缺失型大豆的11S/7S比值在4.65以上,高于目前普遍报道的2.0~3.0;3）致敏蛋白α亚基缺失型大豆必需氨基酸含量接近或高于FAO/WHO标准;两种模式下,α亚基缺失型大豆的5种化学评分及贴进度值都很高,接近标准蛋白。致敏蛋白α亚基缺失型大豆蛋白质氨基酸组分平衡,11S/7S比值更优,营养品质更高。
文摘将鸡催乳素 (PRL)和抑制素 - α亚基 (INB- α)基因编码序列重组为融合基因 ,制备了同时包含这 2种激素基因的融合蛋白。通过 PCR和分子克隆的方法首先将全部粤黄鸡 PRL成熟肽 c DNA克隆到载体 p RSET A的 Bgl 和 Eco R 克隆位点之间 ,获得重组质粒 p PRL- RSET。鸡 INB- α片段经扩增后分别被克隆到质粒 p RSET A和 p PRL- RSET的 Nhe 和 Xho 克隆位点之间 ,获得重组质粒 p INB- RSET和 p INB- PRL。以上重组质粒构建的正确性分别由各特定引物组合扩增的 PCR产物长度、特定限制性内切酶消化各重组质粒所得产物长度以及对各质粒的测序结果得到验证。重组质粒 p PRL- RSET和 p INB- PRL 转化 E.coli BL2 1(DE3)株 ,IPTG诱导后所表达的产物经 SDS- PAGE显示 ,其分别与所预期的重组蛋白分子大小相符。质粒 p PRL - RSET和 p INB- PRL的表达产物和用 Ni- NTA凝胶纯化的 2重组蛋白产物都可与抗鸡 PRL 抗体产生特异的免疫印迹 ,并且表达菌裂解液和相应纯化蛋白的免疫印迹处于同一位置。结果说明 ,试验已成功完成了鸡 PRL、INB-α及 2者融合蛋白的构建。
