华北农学报
華北農學報
화북농학보
ACTA AGRICULTURAE BOREALI-SINICA
2013年
5期
59-65
,共7页
王青云%王润青%方聪燕%侯佩%苏亮%李建平%宋梅芳%杨建平%李雪梅%吴大付
王青雲%王潤青%方聰燕%侯珮%囌亮%李建平%宋梅芳%楊建平%李雪梅%吳大付
왕청운%왕윤청%방총연%후패%소량%리건평%송매방%양건평%리설매%오대부
水稻%蛋白磷酸酶6%过表达载体构建%生物信息学分析
水稻%蛋白燐痠酶6%過錶達載體構建%生物信息學分析
수도%단백린산매6%과표체재체구건%생물신식학분석
Oryza sativa%Protein phosphatase 6%Vector construction for overexpression%Bioinformatics analysis
为克隆水稻蛋白磷酸酶6（PP6）催化亚基基因OsPP6C，并构建该基因的过表达载体以及进行生物信息学分析。采用RT-PCR技术从水稻中花11叶片cDNA中扩增OsPP6C基因全长，并构建与GUS融合的pBI121-OsPP6C-GUS表达载体。通过生物信息学方法分析了OsPP6C蛋白的理化性质、与拟南芥AtPP6C（即AtFyPP）之间的同源性以及不同物种间的系统发育关系，此外，对OsPP6C基因的启动子进行了顺式作用元件分析。结果表明，该基因转录序列包含一个912 bp的开放阅读框，编码303个氨基酸残基，蛋白的分子量约为3．47 kDa，等电点为5．13；具有疏水性，但不存在信号肽，属于非分泌型蛋白。 OsPP6C与拟南芥AtFyPP1和AtFyPP3氨基酸序列一致性分别为93．73％和93．40％，系统进化树显示OsPP6C与小麦TaPP6C的亲缘关系最近，而与其他物种亲缘关系相对较远。对该基因启动子中含有的顺式作用元件分析表明，该启动子中除TATA盒和CAAT盒外，还含有多种参与光应答、激素（ ABA、乙烯、生长素、MeJA和赤霉素等）应答以及低温、热激和干旱等非生物胁迫因子应答的调控元件。为进一步研究水稻Os-PP6C基因的表达特性和功能奠定了基础。
為剋隆水稻蛋白燐痠酶6（PP6）催化亞基基因OsPP6C，併構建該基因的過錶達載體以及進行生物信息學分析。採用RT-PCR技術從水稻中花11葉片cDNA中擴增OsPP6C基因全長，併構建與GUS融閤的pBI121-OsPP6C-GUS錶達載體。通過生物信息學方法分析瞭OsPP6C蛋白的理化性質、與擬南芥AtPP6C（即AtFyPP）之間的同源性以及不同物種間的繫統髮育關繫，此外，對OsPP6C基因的啟動子進行瞭順式作用元件分析。結果錶明，該基因轉錄序列包含一箇912 bp的開放閱讀框，編碼303箇氨基痠殘基，蛋白的分子量約為3．47 kDa，等電點為5．13；具有疏水性，但不存在信號肽，屬于非分泌型蛋白。 OsPP6C與擬南芥AtFyPP1和AtFyPP3氨基痠序列一緻性分彆為93．73％和93．40％，繫統進化樹顯示OsPP6C與小麥TaPP6C的親緣關繫最近，而與其他物種親緣關繫相對較遠。對該基因啟動子中含有的順式作用元件分析錶明，該啟動子中除TATA盒和CAAT盒外，還含有多種參與光應答、激素（ ABA、乙烯、生長素、MeJA和赤黴素等）應答以及低溫、熱激和榦旱等非生物脅迫因子應答的調控元件。為進一步研究水稻Os-PP6C基因的錶達特性和功能奠定瞭基礎。
위극륭수도단백린산매6（PP6）최화아기기인OsPP6C，병구건해기인적과표체재체이급진행생물신식학분석。채용RT-PCR기술종수도중화11협편cDNA중확증OsPP6C기인전장，병구건여GUS융합적pBI121-OsPP6C-GUS표체재체。통과생물신식학방법분석료OsPP6C단백적이화성질、여의남개AtPP6C（즉AtFyPP）지간적동원성이급불동물충간적계통발육관계，차외，대OsPP6C기인적계동자진행료순식작용원건분석。결과표명，해기인전록서렬포함일개912 bp적개방열독광，편마303개안기산잔기，단백적분자량약위3．47 kDa，등전점위5．13；구유소수성，단불존재신호태，속우비분비형단백。 OsPP6C여의남개AtFyPP1화AtFyPP3안기산서렬일치성분별위93．73％화93．40％，계통진화수현시OsPP6C여소맥TaPP6C적친연관계최근，이여기타물충친연관계상대교원。대해기인계동자중함유적순식작용원건분석표명，해계동자중제TATA합화CAAT합외，환함유다충삼여광응답、격소（ ABA、을희、생장소、MeJA화적매소등）응답이급저온、열격화간한등비생물협박인자응답적조공원건。위진일보연구수도Os-PP6C기인적표체특성화공능전정료기출。
The aims of this study are to clone the catalytic subunit gene of rice protein phosphatase 6 (OsPP6C),construct the over-expressing vector of this gene and conduct the bioinformatics analysis .Using the RT-PCR technique,the full length of OsPP6C gene was amplified from the complementary DNA (cDNA)of rice Zhong-hua 11 variety,and it was fused with GUS to form pBI121-OsPP6C-GUS expression vector.Through bioinformatics methods,the physiochemical characteristics of OsPP6C protein,the homology among Arabidopsis PP6Cs(i.e.At-FyPPs)and OsPP6C,and the phylogenetic relationship among different species were analyzed;in addition,the cis-acting elements of the OsPP6C promoter were analyzed .The results show that the transcript sequence of this gene contains an open reading frame of 912 base pairs , this gene encodes a peptide of 303 amino acids , the molecular weight and isoelectric point of this protein is approximately 3.47 kDa and 5.13,respectively;the OsPP6C protein The aims of this study are to clone the catalytic subunit gene of rice protein phosphatase 6 (OsPP6C),construct the over-expressing vector of this gene and conduct the bioinformatics analysis .Using the RT-PCR technique,the full length of OsPP6C gene was amplified from the complementary DNA (cDNA)of rice Zhong-hua 11 variety,and it was fused with GUS to form pBI121-OsPP6C-GUS expression vector.Through bioinformatics methods,the physiochemical characteristics of OsPP6C protein,the homology among Arabidopsis PP6Cs(i.e.At-FyPPs)and OsPP6C,and the phylogenetic relationship among different species were analyzed;in addition,the cis-acting elements of the OsPP6C promoter were analyzed .The results show that the transcript sequence of this gene contains an open reading frame of 912 base pairs , this gene encodes a peptide of 303 amino acids , the molecular weight and isoelectric point of this protein is approximately 3.47 kDa and 5.13,respectively;the OsPP6C protein <br> has the hydrophobic property ,but there is no signal peptide in this protein and it belongs to the non-secretory pro-teins.The amino acid identity of OsPP6C with AtFyPP1 and AtFyPP3 was 93.73%and 93.40%,respectively;the phylogenetic tree shows that OsPP6C has the closest genetic relationship with wheat TaPP 6C.The cis-acting element analysis in the promoter of this gene shows that besides the TATA box and CAAT box ,the promoter also contains multiple light-responsive elements ,various responsive elements in relation to hormones such as ABA ,ethylene ,aux-in,methyl jasmonate ( MeJA ) and gibberellic acid , and the regulatory elements involved in various abiotic stresses such as low temperature ,heat stress and drought .This paper will lay a solid foundation for further studying expres-sion characteristics and functions of OsPP6 C gene in rice .
