A 105-k Da polymer lectin was purified from lamprey（Lampetra japonica） serum by chromatography methods including cation ion-exchange chromatography with a SP-Sepharose TM XL column and size exclusion chromatography ...A 105-k Da polymer lectin was purified from lamprey（Lampetra japonica） serum by chromatography methods including cation ion-exchange chromatography with a SP-Sepharose TM XL column and size exclusion chromatography with a Superdex 200 column. The target fractions were collected according to the direction of hemagglutinating activity. The results revealed that the active fractions could adsorb on SP-Sepharose column and showed a 280 nm UV absorbance peak corresponding to molecular weights of 105 k Da in the following size exclusion chromatography. The target fractions with hemagglutinating activity were further checked by NativePAGE and SDS-PAGE. Two single bands at around 105 k Da and 35 k Da were displayed by two electrophoresis methods respectively, indicating that the protein exists as a trimer in solution. After Native-PAGE and SDS-PAGE,two bands were excised from the gels respectively and further identified by MALDI-TOF/TOF as serum lectin（gi：13094239）. The lectin was able to agglutinate rabbit red blood cells（RRBCs） and sheep red blood cells（SRBCs） in vitro. The lectin isolated from lamprey serum in the current study might be helpful for deeply understanding the innate immune molecules dependent immune defence in jawless vertebrates which have been proved recently that they possess a lymphocyte-based system of anticipatory immunity with variable lymphocyte receptors as mediators.展开更多
为了研究桉树不同无性系在铝处理下的叶片蛋白表达的差异,并从分子生物学角度预测与阐释桉树耐铝性机制,以巨尾桉(Eucalyptus grandis×E.urophylla)9号、巨尾桉(E.grandis×E.urophylla)12号、韦赤桉(E.wetarensis×E.cama...为了研究桉树不同无性系在铝处理下的叶片蛋白表达的差异,并从分子生物学角度预测与阐释桉树耐铝性机制,以巨尾桉(Eucalyptus grandis×E.urophylla)9号、巨尾桉(E.grandis×E.urophylla)12号、韦赤桉(E.wetarensis×E.camaldulensis)3号和尾叶桉(E.urophylla)4号苗木为材料,并对其进行铝处理。结果表明:从4种无性系叶所得蛋白有明显的上调或下调表达。通过丰度计算,共得蛋白点(P<0.05)98个,巨尾桉9号为14个、巨尾桉12号为38个、尾叶桉4号为33个、韦赤桉3号为13个。经质谱分析,有11个蛋白成功鉴定,包括1个Ru Bis CO、3个Ru Bis CO LSU、1个放氧增强蛋白、1个叶绿素a/b结合蛋白、3个2-半胱氨酸过氧化物酶、1个质体谷氨酰胺合成酶、1个3-磷酸甘油醛脱氢酶,其主要功能与植物自身N和能量代谢以及抗氧化和光合作用等有关。4种不同的桉树无性系应对铝胁迫时有不同的应答机制,差异蛋白质呈现不同的表达量,并且从该角度分析耐铝性最强的是巨尾桉9号,而较弱的是尾叶桉4号。展开更多
基金The National Program on Key Basic Research Project(973 Program)of China under contract No.2013CB835304the National Marine Public Projects under contract No.201305016+1 种基金the National Natural Science Foundation of China under contract Nos31772884 and 31601865the Key Projects of Scientific Research Platform of Liaoning Provincial Education Department under contract No.L201683651
文摘A 105-k Da polymer lectin was purified from lamprey（Lampetra japonica） serum by chromatography methods including cation ion-exchange chromatography with a SP-Sepharose TM XL column and size exclusion chromatography with a Superdex 200 column. The target fractions were collected according to the direction of hemagglutinating activity. The results revealed that the active fractions could adsorb on SP-Sepharose column and showed a 280 nm UV absorbance peak corresponding to molecular weights of 105 k Da in the following size exclusion chromatography. The target fractions with hemagglutinating activity were further checked by NativePAGE and SDS-PAGE. Two single bands at around 105 k Da and 35 k Da were displayed by two electrophoresis methods respectively, indicating that the protein exists as a trimer in solution. After Native-PAGE and SDS-PAGE,two bands were excised from the gels respectively and further identified by MALDI-TOF/TOF as serum lectin（gi：13094239）. The lectin was able to agglutinate rabbit red blood cells（RRBCs） and sheep red blood cells（SRBCs） in vitro. The lectin isolated from lamprey serum in the current study might be helpful for deeply understanding the innate immune molecules dependent immune defence in jawless vertebrates which have been proved recently that they possess a lymphocyte-based system of anticipatory immunity with variable lymphocyte receptors as mediators.
文摘为了研究桉树不同无性系在铝处理下的叶片蛋白表达的差异,并从分子生物学角度预测与阐释桉树耐铝性机制,以巨尾桉(Eucalyptus grandis×E.urophylla)9号、巨尾桉(E.grandis×E.urophylla)12号、韦赤桉(E.wetarensis×E.camaldulensis)3号和尾叶桉(E.urophylla)4号苗木为材料,并对其进行铝处理。结果表明:从4种无性系叶所得蛋白有明显的上调或下调表达。通过丰度计算,共得蛋白点(P<0.05)98个,巨尾桉9号为14个、巨尾桉12号为38个、尾叶桉4号为33个、韦赤桉3号为13个。经质谱分析,有11个蛋白成功鉴定,包括1个Ru Bis CO、3个Ru Bis CO LSU、1个放氧增强蛋白、1个叶绿素a/b结合蛋白、3个2-半胱氨酸过氧化物酶、1个质体谷氨酰胺合成酶、1个3-磷酸甘油醛脱氢酶,其主要功能与植物自身N和能量代谢以及抗氧化和光合作用等有关。4种不同的桉树无性系应对铝胁迫时有不同的应答机制,差异蛋白质呈现不同的表达量,并且从该角度分析耐铝性最强的是巨尾桉9号,而较弱的是尾叶桉4号。
