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不同失水状态发菜类囊体膜蛋白的分离及鉴定
  • ISSN号:1000-4025
  • 期刊名称:《西北植物学报》
  • 时间:0
  • 分类:Q503[生物学—生物化学]
  • 作者机构:[1]宁夏大学生命科学学院,银川750021
  • 相关基金:国家自然科学基金(31360054,31060038)
中文摘要:

采用液氮研磨与超声波破碎相结合的方法,破碎不同失水状态的发菜藻体和细胞,用差速离心法制备发菜类囊体膜粗制品,蔗糖密度梯度高速离心纯化类囊体膜,SDS-PAGE电泳分离类囊体膜蛋白,并对膜蛋白进行了MALDI-TOF-TOF/MS质谱分析和鉴定。结果表明:(1)充分吸胀4 h后失水6 h的发菜(含水量51.2%)和失水24 h的发菜(含水量14.9%),经过多步差速离心后再进行蔗糖密度梯度高速离心,可得到纯化的类囊体膜。(2)发菜类囊体膜蛋白SDS-PAGE电泳分离到14个条带,共鉴定出8种蛋白,根据其功能可分为4类——光合作用相关蛋白(光系统Ⅱ锰稳定蛋白PsbO,F1F0 ATP合成酶α亚基和β亚基)、结构域蛋白、选择性通道蛋白OprB、未知蛋白(hypothetical protein Npun_R1321、Npun_R3785、N9414_02186),它们在发菜的光合作用中具有重要作用。

英文摘要:

The method of liquid nitrogen grinding and ultrasonic were applied to crush Nostoc flagelliforme colonies and cells.The crudes were isolated by differential centrifugation;thylakoid membranes were purified by high speed centrifugation with gradient density sucrose;the thylakoid membrane proteins were extracted and separated by SDS-PAGE.The results suggested that:(1)Thylakoid membrane of colonies on dehydration for 6 h(water contenting 51.2%) and on dehydration for 24 h(water contenting 14.9%) were purified by many tims differential centrifugation and then by high speed centrifugation with gradient density sucrose.(2)14 bands of thylakoid membrane protein were separated by SDS-PAGE,their mass weight mainly ranged from 25 kD to 60 kD.8 kinds of protein were identified by MALDI-TOF-TOF/MS and database searching.According to their biological functions,these thylakoid membrane proteins were divided into 4 categories,including photosynthesis proteins(photosystem Ⅱ manganese-stabilizing protein PsbO,F1F0 ATP synthase subunit alpha,F1F0 ATP synthase subunit beta),binding domain protein(hemerythrin HHE cation binding domain-containing protein),carbohydrate-selective porin(carbohydrate-selective porin OprB),unknown proteins(hypothetical protein Npun_R1321,Npun_R3785 and N9414_02186).They might play an important role in photosynthesis of N.flagelliforme.

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期刊信息
  • 《西北植物学报》
  • 北大核心期刊(2011版)
  • 主管单位:中华人民共和国教育部
  • 主办单位:西北农林科技大学 陕西省植物学会
  • 主编:赵忠
  • 地址:陕西杨陵邰城路3号西北农林科技大学
  • 邮编:712100
  • 邮箱:xbzwxb@vip.163.com
  • 电话:029-87082936
  • 国际标准刊号:ISSN:1000-4025
  • 国内统一刊号:ISSN:61-1091/Q
  • 邮发代号:52-73
  • 获奖情况:
  • 全国优秀期刊,中国自然科学核心期刊,中国期刊方阵“双效”期刊
  • 国内外数据库收录:
  • 俄罗斯文摘杂志,美国化学文摘(网络版),英国农业与生物科学研究中心文摘,美国生物科学数据库,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:46104