激光生物学报
激光生物學報
격광생물학보
ACTA LASER BIOLOGY SINICA
2011年
1期
61-66
,共6页
刘振宇%吴祖建%谢荔岩%林奇英%谢联辉
劉振宇%吳祖建%謝荔巖%林奇英%謝聯輝
류진우%오조건%사려암%림기영%사련휘
质体蓝素基因%孔石莼%RACE%cDNA
質體藍素基因%孔石莼%RACE%cDNA
질체람소기인%공석순%RACE%cDNA
plastocyanin gene%Ulva pertusa%RACE%cDNA
采用cDNA末端快速扩增的办法,从孔石莼(Ulva pertusa)中克隆获得质体蓝素基因.该基因完整的cDNA为787 bp,包括40 bp 5'端非编码区和327 bp的3'端非编码区,以及一个420 bp的开放阅读框架,编码139个氨基酸的蛋白质.该基因编码质体蓝素的前体肽,其N端41个氨基酸残基为信号肽,后面为98个氨基酸残基的成熟肽.从Genbank中选择了13个质体蓝素的前体肽基因进行序列比对分析和构建进化树.孔石莼质体蓝素基因与其它质体蓝素基因的同源性为48.2%至78.8%.该进化树将来源于6种藻类植物的7个质体蓝素基因聚类在一起,显示出它们较近的进化关系.同样,也表现出11种生物的分子进化关系.序列比对结果显示,在质体蓝素的基因序列中存在两个高度保守的基序,它编码质体蓝素蛋白的铜结合活性位点.
採用cDNA末耑快速擴增的辦法,從孔石莼(Ulva pertusa)中剋隆穫得質體藍素基因.該基因完整的cDNA為787 bp,包括40 bp 5'耑非編碼區和327 bp的3'耑非編碼區,以及一箇420 bp的開放閱讀框架,編碼139箇氨基痠的蛋白質.該基因編碼質體藍素的前體肽,其N耑41箇氨基痠殘基為信號肽,後麵為98箇氨基痠殘基的成熟肽.從Genbank中選擇瞭13箇質體藍素的前體肽基因進行序列比對分析和構建進化樹.孔石莼質體藍素基因與其它質體藍素基因的同源性為48.2%至78.8%.該進化樹將來源于6種藻類植物的7箇質體藍素基因聚類在一起,顯示齣它們較近的進化關繫.同樣,也錶現齣11種生物的分子進化關繫.序列比對結果顯示,在質體藍素的基因序列中存在兩箇高度保守的基序,它編碼質體藍素蛋白的銅結閤活性位點.
채용cDNA말단쾌속확증적판법,종공석순(Ulva pertusa)중극륭획득질체람소기인.해기인완정적cDNA위787 bp,포괄40 bp 5'단비편마구화327 bp적3'단비편마구,이급일개420 bp적개방열독광가,편마139개안기산적단백질.해기인편마질체람소적전체태,기N단41개안기산잔기위신호태,후면위98개안기산잔기적성숙태.종Genbank중선택료13개질체람소적전체태기인진행서렬비대분석화구건진화수.공석순질체람소기인여기타질체람소기인적동원성위48.2%지78.8%.해진화수장래원우6충조류식물적7개질체람소기인취류재일기,현시출타문교근적진화관계.동양,야표현출11충생물적분자진화관계.서렬비대결과현시,재질체람소적기인서렬중존재량개고도보수적기서,타편마질체람소단백적동결합활성위점.
A plastocyanin precursor cDNA gene of Ulva pertusa was isolated with the method of rapid amplification of cDNA ends (RACE). The complete cDNA sequence covered 787 bp nucleotides including a 420 bp of open reading frame which encodes a putative plastocyanin precursor of 139 amino acid residues, a 40 bp 5' - terminal and a 327 bp 3'-terminal untranslated regions. The cDNA gene from U. pertusa shared homology from 48.2 % to 78.8 % with the 12genes from 10 different species. A phylogenetic tree was constructed based on the 13 sequences of plastocyanin precursor genes from 11 species. 7 plastocyanin gene sequences from 6 algae formed a cluster in the phylogenetic tree, indicating closely evolutionary relationship. The phylogenetic tree also revealed the molecular evolutionary relationship of the 11species based on the 13 sequences analyses. Sequence alignments also represented two highly conserved motifs in the sequences, which encoded the copper-binding active sites of the plastocyanin protein.