CN114621933B - 一种accD突变蛋白及其应用 - Google Patents
一种accD突变蛋白及其应用 Download PDFInfo
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Abstract
本发明公开了一种accD突变蛋白及其应用,所述accD突变蛋白的氨基酸序列如SEQ ID NO.3所示。本发明通过将accD突变蛋白在植物中过表达,使得植物在高温下的育性显著增强,同时由于叶绿体基因在植物中具有很高的保守性,因此本发明的accD突变蛋白具有普适性,可以用于各种植物中,解决因高温导致的植物雄性败育以及由此引起的大幅减产问题。
Description
技术领域
本发明属于基因工程技术领域,具体涉及一种accD突变蛋白及其应用。
背景技术
植物雄性不育是一个与农业生产密切相关的植物学性状,是植物在发育过程中基因型表达与环境相互作用的结果。植株本身的雄性不育就可以在免去人工去雄的情况下作为遗传工具用于开发利用作物杂种优势,进行轮回选择,回交等育种研究。利用植物的雄性不育性培育各种雄性不育系,再借助遗传工程大量生产杂交种子,从而使许多作物特别是自花传粉作物的杂种优势得以在生产上利用,为大规模生产杂交种子提供了可能性。
相关技术主要针对的常温下提高植物育性或者恢复植物育性的方法,而能够显著增强高温下育性的方法很少;且绝大多数育性相关的技术都是开发的与育性相关的控制基因、分子标记等,这些都是基于该植物本身特有,其保守性有待考究,在其他植物中的应用前景不明;而通过杂交等方法提高育性的方法繁杂。通过外施植物生长调节剂提高育性等方法,对于使用量的把握需要精确,不利于广泛使用。
温度是影响植物生长发育的重要因素,高温尤其影响花粉发育,从而导致严重影响植物的育性,降低植物产量。分子生物学手段是改善植物高温耐受,提高植物育性的重要手段之一。
发明内容
本发明旨在至少解决上述现有技术中存在的技术问题之一。为此,本发明提出一种蛋白质,能够有效的提高植物高温下的育性。
本发明还提出一种编码上述蛋白质的核酸分子。
本发明还提出一种与上述核酸分子相关的生物材料。
本发明还提出一种用于扩增上述核酸分子的引物。
本发明还提出上述蛋白质、编码上述蛋白质的核酸分子、生物材料、引物的应用。
本发明还提出一种抗逆植物的培育方法。
本发明还提出一种促进植物生长的方法。
在本发明的一个方面,提出了一种蛋白质,是如下a)或b)或c)或d)的蛋白质:
a)氨基酸序列如SEQ ID NO.3所示的蛋白质;
b)氨基酸序列如SEQ ID NO.3所示的蛋白质的N端和/或C端连接标签得到的融合蛋白质;
c)将序列如SEQ ID NO.3所示的氨基酸序列经过一个或几个氨基酸残基的取代和/或缺失和/或添加得到的具有相同功能的蛋白质;
d)与序列如SEQ ID NO.3所示的氨基酸序列具有75%或75%以上的同源性且具有相同功能的蛋白质。
在本发明的一些实施方式中,所述蛋白质为accD突变蛋白。
在本发明的第二方面,提出了一种编码上述蛋白质的核酸分子。
在本发明的一些实施方式中,核酸分子为accD突变基因。
在本发明的一些实施方式中,所述核酸分子的核苷酸序列如SEQ ID NO.1所示。
在本发明的第三方面,提出了一种与上述核酸分子相关的生物材料,为下述1)至7)中的任一种:
1)含有上述核酸分子的表达盒;
2)含有上述核酸分子的重组载体;
3)含有1)所述表达盒的重组载体;
4)含有上述核酸分子的重组微生物;
5)含有1)所述表达盒的重组微生物;
6)含有2)所述重组载体的重组微生物;
7)含有3)所述重组载体的重组微生物。
一种重组载体,所述载体包括上述核酸分子和前导肽序列。
在本发明的一些实施方式中,所述前导肽序列如SEQ ID NO.7所示。
在本发明的第四方面,提出了一种用于扩增上述核酸分子的引物。
在本发明的一些实施方式中,所述引物包括上游引物序列和下游引物序列,所述上游引物核苷酸序列如SEQ ID NO.5所示,下游引物序列如SEQ ID NO.6所示。
在本发明的第五方面,提出了上述蛋白质、编码上述蛋白质的核酸分子、生物材料、引物的应用,所述应用为在植物育种中的应用。
在本发明的一些实施方式中,所述应用为抗逆植物辅助育种中的应用,所述抗逆为抗高温。
在本发明的一些实施方式中,所述应用为提高高温下转基因植物育性中的应用。
在本发明的一些实施方式中,所述高温为温度为28-37℃。
在本发明的第六方面,提出了一种抗逆植物的培育方法,包括将将上述核酸分子、生物材料转入受体植物中,得到转基因植物。
在本发明的一些实施方式中,所述植物为拟南芥、油菜或棉花。
在本发明的第七方面,提出了一种促进植物生长的方法,所述方法包括如下步骤:将上述核酸分子、生物材料转入受体植物中。
根据本发明的实施方式,至少具有以下有益效果:本发明通过将accD蛋白第265号位置的丝氨酸替换成亮氨酸,得到accD突变蛋白,将accD突变蛋白在植物中过表达,使得植物在高温下的育性显著增强,同时由于叶绿体基因在植物中具有很高的保守性,因此本发明的accD突变蛋白具有普适性,可以用于各种植物中,解决因高温导致的植物雄性败育以及由此引起的大幅减产问题。
附图说明
下面结合附图和实施例对本发明做进一步的说明,其中:
图1为本发明实施例1中的accD突变基因的制备流程;
图2为本发明测试例中的转基因植株western blot检测结果图,其中OE-1、OE-5、OE-12为转基因植株;col为野生型的植株;
图3为本发明测试例中的accD突变基因对高温下的拟南芥的生长结果图,其中,其中OE-1、OE-5、OE-12为转基因植株;col为野生型的植株。
具体实施方式
以下将结合实施例对本发明的构思及产生的技术效果进行清楚、完整地描述,以充分地理解本发明的目的、特征和效果。显然,所描述的实施例只是本发明的一部分实施例,而不是全部实施例,基于本发明的实施例,本领域的技术人员在不付出创造性劳动的前提下所获得的其他实施例,均属于本发明保护的范围。
实施例1
本实施例提供了一种accD突变蛋白,技术方案如图1所示,具体为通过生物信息学分析得到在植物中保守的基因accD,有选择的将第794号位置的胞嘧啶C替换成胸腺嘧啶T,accD基因如SEQ ID NO.2所示,accD突变基因如SEQ ID NO.1所示,得到的accD突变蛋白的氨基酸序列如SEQ ID NO.3所示,accD蛋白原始氨基酸序列如SEQ ID NO.4所示,accD蛋白与accD突变蛋白的区别在于,将accD蛋白第265号位置的丝氨酸替换成亮氨酸。
accD突变基因核苷酸序列:
ATGGAAAAATCGTGGTTCAATTTTATGTTTTCTAAGGGAGAATTGGAATACAGAGGTGAGCTAAGTAAAGCAATGGATAGTTTTGCTCCTGGTGAAAAGACTACTATAAGTCAAGACCGTTTTATATATGATATGGATAAAAACTTTTATGGTTGGGATGAGCGTTCTAGTTATTCTTCTAGTTATTCCAATAATGTTGATCTTTTAGTTAGCTCCAAGGACATTCGCAATTTCATATCGGATGACACCTTTTTTGTTAGGGATAGTAATAAGAATAGTTATTCTATATTTTTTGATAAAAAAAAAAAAATTTTTGAGATTGACAATGATTTTAGTGACCTAGAAAAATTTTTTTATAGTTATTGTAGTTCTAGTTATCTAAATAATAGATCTAAAGGTGACAACGATCTGCACTATGATCCTTACATTAAGGATACTAAATATAATTGTACTAATCACATTAATAGTTGCATTGATTCTTATTTTCGTTCTTACATCTGTATTGATAATAACTTTTTAATCGATAGTAATAATTTTAATGAAAGTTACATTTATAATTTCATTTGTAGTGAAAGCGGAAAGATTCGTGAAAGTAAAAATTACAAGATAAGAACTAATAGGAATCGTAGTAATTTAATAAGTTCTAAGGATTTCGATATAACTCAAAACTACAATCAATTGTGGATTCAATGCGACAATTGTTATGGATTAATGTATAAGAAAGTCAAAATGAATGTTTGTGAACAATGTGGACATTATTTGAAAATGAGTAGTTCAGAAAGAATCGAGCTTTTGATTGATCCGGGTACTTGGAATCCTATGGATGAAGACATGGTCTCTGCGGATCCCATTAAATTTCATTCGAAGGAGGAACCTTATAAAAACCGTATTGACTCTGCGCAAAAAACTACAGGATTGACTGACGCTGTTCAAACAGGTACAGGTCAACTAAACGGTATTCCGGTAGCCCTTGGGGTTATGGATTTTCGGTTTATGGGGGGTAGTATGGGATCCGTAGTAGGCGAAAAAATAACTCGTTTGATCGAGTATGCTACCAATCAATGTTTACCTCTTATTTTAGTGTGTTCTTCCGGAGGAGCACGAATGCAAGAAGGAAGTTTAAGTTTGATGCAAATGGCTAAAATTTCTTCGGTTTTATGTGATTATCAATCAAGTAAAAAGTTATTCTATATATCAATTCTTACATCTCCTACTACCGGTGGAGTGACAGCTAGTTTTGGTATGTTGGGGGATATCATTATTGCCGAACCCTATGCCTATATTGCATTTGCGGGTAAAAGAGTAATTGAACAAACATTGAAAAAAGCCGTGCCTGAAGGTTCACAAGCGGCTGAATCTTTATTACGTAAGGGCTTATTGGATGCAATTGTACCACGTAATCTTTTAAAAGGTGTTCTGAGCGAGTTATTTCAGCTCCATGCTTTTTTTCCTTTGAACACAAAT(SEQ ID NO.1)。
accD突变蛋白氨基酸序列:
MEKSWFNFMFSKGELEYRGELSKAMDSFAPGEKTTISQDRFIYDMDKNFYGWDERSSYSSSYSNNVDLLVSSKDIRNFISDDTFFVRDSNKNSYSIFFDKKKKIFEIDNDFSDLEKFFYSYCSSSYLNNRSKGDNDLHYDPYIKDTKYNCTNHINSCIDSYFRSYICIDNNFLIDSNNFNESYIYNFICSESGKIRESKNYKIRTNRNRSNLISSKDFDITQNYNQLWIQCDNCYGLMYKKVKMNVCEQCGHYLKMSSSERIELLIDPGTWNPMDEDMVSADPIKFHSKEEPYKNRIDSAQKTTGLTDAVQTGTGQLNGIPVALGVMDFRFMGGSMGSVVGEKITRLIEYATNQCLPLILVCSSGGARMQEGSLSLMQMAKISSVLCDYQSSKKLFYISILTSPTTGGVTASFGMLGDIIIAEPYAYIAFAGKRVIEQTLKKAVPEGSQAAESLLRKGLLDAIVPRNLLKGVLSELFQLHAFFPLNTN(SEQ ID NO.3)。
实施例2载体的制备
融合载体的制备包括以下步骤:
(1)利用Xba1和Pst1限制性内切酶(购自上海生工生物工程有限公司)双酶切pCAMBIA1300(融合有35S强启动子)载体(购自上海生工生物工程有限公司);双酶切体系如表1所示;反应条件:37℃,4h。
表1质粒的双酶切体系
将经双酶切后的质粒进行脂糖凝胶电泳检测,采用胶回收试剂盒(TaKaRaMiniBEST Agarose Gel DNA Extraction Kit)进行回收目的条带,采用分光光度计测定回收后质粒浓度。
(2)利用accD突变基因扩增引物序列(序列SEQ ID NO.5~6),引物序列如表2所示,以拟南芥cDNA为模板,扩增accD突变基因序列,将扩增得到的accD突变基因序列利用Xba1和Pst1两个酶切位点进行双酶切,与双酶切的pCAMBIA1300质粒载体采用T4DNA连接酶(购自上海生工生物工程有限公司)进行连接,得到重组载体pCAMBIA1300-accD。
表2引物序列
(3)利用Sac1和Xba1两个酶切位点,双酶切pCAMBIA1300-accD载体;双酶切体系如表1所示。
(4)利用PORA前导肽扩增引物序列(序列如SEQ ID NO.8-9所示),以拟南芥cDNA为模板,扩增PORA前导肽序列(序列如SEQ ID NO.7所示),将扩增得到的PORA前导肽序列利用Sac1和Xba1两个酶切位点进行双酶切,与双酶切的pCAMBIA1300-accD载体采用T4DNA连接酶(购自上海生工生物工程有限公司)进行连接,得到重组载体,将重组载体进行测序确认。
实施例2转基因植株的获得
1、重组质粒的转化
将实施例构建好的重组载体电击转化农杆菌菌株(购自天根生化),得到重组农杆菌,在kan和Rif抗性培养基上培养两天后,挑取单菌落,摇菌鉴定阳性克隆。
2、菌液的准备
将鉴定为阳性的农杆菌感受态细胞在28℃过夜培养,取5-6ml菌液于4000rpm离心5min后,去上清,使用含MES(10mM),MgCl(10mM),AS(150uM)的重悬液重悬至OD600值为0.8左右,28℃避光静置2~3h左右,得到侵染菌液。
3、侵染
通过花浸泡法(花浸在步骤2的菌液中30秒)将重组质粒分别导入60株拟南芥,对转化后的拟南芥进行种子收集,干燥一周后利用潮霉素进行抗性筛选,筛选出具有抗性的阳性苗。
4、筛选获得转基因植株
随机选取3株阳性苗进行Western blot检测,检测结果如图2所示,从图2中可以看出,3株随机检测的转基因植株中accD突变蛋白都成功实现了过表达。
测试例转基因植物的高温育性分析
实验组采用实施例2获得的转基因拟南芥,对照组采用野生型的拟南芥,每组20株,将对照组和实验组的拟南芥温度采用30℃进行培养,其他参数均为常规方法。在高温培养21天和26天后分别进行拍照记录。在结果后,统计实验组和对照组的果荚平均数。
表3
组别 | 果荚数(个/株) |
实验组 | 1 |
对照组 | 8 |
实验结果如图3和表3所示,从图中可以看出,高温处理21天和26天后,accD突变基因可以有效的促进拟南芥的生长,从表3中可以看出,accD突变基因可以有效的提高高温下拟南芥的结实率。
上面结合附图对本发明实施例作了详细说明,但是本发明不限于上述实施例,在所属技术领域普通技术人员所具备的知识范围内,还可以在不脱离本发明宗旨的前提下作出各种变化。此外,在不冲突的情况下,本发明的实施例及实施例中的特征可以相互组合。
序列表
<110> 湖南农业大学
<120> 一种accD突变蛋白及其应用
<160> 9
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1464
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
atggaaaaat cgtggttcaa ttttatgttt tctaagggag aattggaata cagaggtgag 60
ctaagtaaag caatggatag ttttgctcct ggtgaaaaga ctactataag tcaagaccgt 120
tttatatatg atatggataa aaacttttat ggttgggatg agcgttctag ttattcttct 180
agttattcca ataatgttga tcttttagtt agctccaagg acattcgcaa tttcatatcg 240
gatgacacct tttttgttag ggatagtaat aagaatagtt attctatatt ttttgataaa 300
aaaaaaaaaa tttttgagat tgacaatgat tttagtgacc tagaaaaatt tttttatagt 360
tattgtagtt ctagttatct aaataataga tctaaaggtg acaacgatct gcactatgat 420
ccttacatta aggatactaa atataattgt actaatcaca ttaatagttg cattgattct 480
tattttcgtt cttacatctg tattgataat aactttttaa tcgatagtaa taattttaat 540
gaaagttaca tttataattt catttgtagt gaaagcggaa agattcgtga aagtaaaaat 600
tacaagataa gaactaatag gaatcgtagt aatttaataa gttctaagga tttcgatata 660
actcaaaact acaatcaatt gtggattcaa tgcgacaatt gttatggatt aatgtataag 720
aaagtcaaaa tgaatgtttg tgaacaatgt ggacattatt tgaaaatgag tagttcagaa 780
agaatcgagc ttttgattga tccgggtact tggaatccta tggatgaaga catggtctct 840
gcggatccca ttaaatttca ttcgaaggag gaaccttata aaaaccgtat tgactctgcg 900
caaaaaacta caggattgac tgacgctgtt caaacaggta caggtcaact aaacggtatt 960
ccggtagccc ttggggttat ggattttcgg tttatggggg gtagtatggg atccgtagta 1020
ggcgaaaaaa taactcgttt gatcgagtat gctaccaatc aatgtttacc tcttatttta 1080
gtgtgttctt ccggaggagc acgaatgcaa gaaggaagtt taagtttgat gcaaatggct 1140
aaaatttctt cggttttatg tgattatcaa tcaagtaaaa agttattcta tatatcaatt 1200
cttacatctc ctactaccgg tggagtgaca gctagttttg gtatgttggg ggatatcatt 1260
attgccgaac cctatgccta tattgcattt gcgggtaaaa gagtaattga acaaacattg 1320
aaaaaagccg tgcctgaagg ttcacaagcg gctgaatctt tattacgtaa gggcttattg 1380
gatgcaattg taccacgtaa tcttttaaaa ggtgttctga gcgagttatt tcagctccat 1440
gctttttttc ctttgaacac aaat 1464
<210> 2
<211> 1464
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
atggaaaaat cgtggttcaa ttttatgttt tctaagggag aattggaata cagaggtgag 60
ctaagtaaag caatggatag ttttgctcct ggtgaaaaga ctactataag tcaagaccgt 120
tttatatatg atatggataa aaacttttat ggttgggatg agcgttctag ttattcttct 180
agttattcca ataatgttga tcttttagtt agctccaagg acattcgcaa tttcatatcg 240
gatgacacct tttttgttag ggatagtaat aagaatagtt attctatatt ttttgataaa 300
aaaaaaaaaa tttttgagat tgacaatgat tttagtgacc tagaaaaatt tttttatagt 360
tattgtagtt ctagttatct aaataataga tctaaaggtg acaacgatct gcactatgat 420
ccttacatta aggatactaa atataattgt actaatcaca ttaatagttg cattgattct 480
tattttcgtt cttacatctg tattgataat aactttttaa tcgatagtaa taattttaat 540
gaaagttaca tttataattt catttgtagt gaaagcggaa agattcgtga aagtaaaaat 600
tacaagataa gaactaatag gaatcgtagt aatttaataa gttctaagga tttcgatata 660
actcaaaact acaatcaatt gtggattcaa tgcgacaatt gttatggatt aatgtataag 720
aaagtcaaaa tgaatgtttg tgaacaatgt ggacattatt tgaaaatgag tagttcagaa 780
agaatcgagc tttcgattga tccgggtact tggaatccta tggatgaaga catggtctct 840
gcggatccca ttaaatttca ttcgaaggag gaaccttata aaaaccgtat tgactctgcg 900
caaaaaacta caggattgac tgacgctgtt caaacaggta caggtcaact aaacggtatt 960
ccggtagccc ttggggttat ggattttcgg tttatggggg gtagtatggg atccgtagta 1020
ggcgaaaaaa taactcgttt gatcgagtat gctaccaatc aatgtttacc tcttatttta 1080
gtgtgttctt ccggaggagc acgaatgcaa gaaggaagtt taagtttgat gcaaatggct 1140
aaaatttctt cggttttatg tgattatcaa tcaagtaaaa agttattcta tatatcaatt 1200
cttacatctc ctactaccgg tggagtgaca gctagttttg gtatgttggg ggatatcatt 1260
attgccgaac cctatgccta tattgcattt gcgggtaaaa gagtaattga acaaacattg 1320
aaaaaagccg tgcctgaagg ttcacaagcg gctgaatctt tattacgtaa gggcttattg 1380
gatgcaattg taccacgtaa tcttttaaaa ggtgttctga gcgagttatt tcagctccat 1440
gctttttttc ctttgaacac aaat 1464
<210> 3
<211> 488
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Met Glu Lys Ser Trp Phe Asn Phe Met Phe Ser Lys Gly Glu Leu Glu
1 5 10 15
Tyr Arg Gly Glu Leu Ser Lys Ala Met Asp Ser Phe Ala Pro Gly Glu
20 25 30
Lys Thr Thr Ile Ser Gln Asp Arg Phe Ile Tyr Asp Met Asp Lys Asn
35 40 45
Phe Tyr Gly Trp Asp Glu Arg Ser Ser Tyr Ser Ser Ser Tyr Ser Asn
50 55 60
Asn Val Asp Leu Leu Val Ser Ser Lys Asp Ile Arg Asn Phe Ile Ser
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Asp Asp Thr Phe Phe Val Arg Asp Ser Asn Lys Asn Ser Tyr Ser Ile
85 90 95
Phe Phe Asp Lys Lys Lys Lys Ile Phe Glu Ile Asp Asn Asp Phe Ser
100 105 110
Asp Leu Glu Lys Phe Phe Tyr Ser Tyr Cys Ser Ser Ser Tyr Leu Asn
115 120 125
Asn Arg Ser Lys Gly Asp Asn Asp Leu His Tyr Asp Pro Tyr Ile Lys
130 135 140
Asp Thr Lys Tyr Asn Cys Thr Asn His Ile Asn Ser Cys Ile Asp Ser
145 150 155 160
Tyr Phe Arg Ser Tyr Ile Cys Ile Asp Asn Asn Phe Leu Ile Asp Ser
165 170 175
Asn Asn Phe Asn Glu Ser Tyr Ile Tyr Asn Phe Ile Cys Ser Glu Ser
180 185 190
Gly Lys Ile Arg Glu Ser Lys Asn Tyr Lys Ile Arg Thr Asn Arg Asn
195 200 205
Arg Ser Asn Leu Ile Ser Ser Lys Asp Phe Asp Ile Thr Gln Asn Tyr
210 215 220
Asn Gln Leu Trp Ile Gln Cys Asp Asn Cys Tyr Gly Leu Met Tyr Lys
225 230 235 240
Lys Val Lys Met Asn Val Cys Glu Gln Cys Gly His Tyr Leu Lys Met
245 250 255
Ser Ser Ser Glu Arg Ile Glu Leu Leu Ile Asp Pro Gly Thr Trp Asn
260 265 270
Pro Met Asp Glu Asp Met Val Ser Ala Asp Pro Ile Lys Phe His Ser
275 280 285
Lys Glu Glu Pro Tyr Lys Asn Arg Ile Asp Ser Ala Gln Lys Thr Thr
290 295 300
Gly Leu Thr Asp Ala Val Gln Thr Gly Thr Gly Gln Leu Asn Gly Ile
305 310 315 320
Pro Val Ala Leu Gly Val Met Asp Phe Arg Phe Met Gly Gly Ser Met
325 330 335
Gly Ser Val Val Gly Glu Lys Ile Thr Arg Leu Ile Glu Tyr Ala Thr
340 345 350
Asn Gln Cys Leu Pro Leu Ile Leu Val Cys Ser Ser Gly Gly Ala Arg
355 360 365
Met Gln Glu Gly Ser Leu Ser Leu Met Gln Met Ala Lys Ile Ser Ser
370 375 380
Val Leu Cys Asp Tyr Gln Ser Ser Lys Lys Leu Phe Tyr Ile Ser Ile
385 390 395 400
Leu Thr Ser Pro Thr Thr Gly Gly Val Thr Ala Ser Phe Gly Met Leu
405 410 415
Gly Asp Ile Ile Ile Ala Glu Pro Tyr Ala Tyr Ile Ala Phe Ala Gly
420 425 430
Lys Arg Val Ile Glu Gln Thr Leu Lys Lys Ala Val Pro Glu Gly Ser
435 440 445
Gln Ala Ala Glu Ser Leu Leu Arg Lys Gly Leu Leu Asp Ala Ile Val
450 455 460
Pro Arg Asn Leu Leu Lys Gly Val Leu Ser Glu Leu Phe Gln Leu His
465 470 475 480
Ala Phe Phe Pro Leu Asn Thr Asn
485
<210> 4
<211> 488
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 4
Met Glu Lys Ser Trp Phe Asn Phe Met Phe Ser Lys Gly Glu Leu Glu
1 5 10 15
Tyr Arg Gly Glu Leu Ser Lys Ala Met Asp Ser Phe Ala Pro Gly Glu
20 25 30
Lys Thr Thr Ile Ser Gln Asp Arg Phe Ile Tyr Asp Met Asp Lys Asn
35 40 45
Phe Tyr Gly Trp Asp Glu Arg Ser Ser Tyr Ser Ser Ser Tyr Ser Asn
50 55 60
Asn Val Asp Leu Leu Val Ser Ser Lys Asp Ile Arg Asn Phe Ile Ser
65 70 75 80
Asp Asp Thr Phe Phe Val Arg Asp Ser Asn Lys Asn Ser Tyr Ser Ile
85 90 95
Phe Phe Asp Lys Lys Lys Lys Ile Phe Glu Ile Asp Asn Asp Phe Ser
100 105 110
Asp Leu Glu Lys Phe Phe Tyr Ser Tyr Cys Ser Ser Ser Tyr Leu Asn
115 120 125
Asn Arg Ser Lys Gly Asp Asn Asp Leu His Tyr Asp Pro Tyr Ile Lys
130 135 140
Asp Thr Lys Tyr Asn Cys Thr Asn His Ile Asn Ser Cys Ile Asp Ser
145 150 155 160
Tyr Phe Arg Ser Tyr Ile Cys Ile Asp Asn Asn Phe Leu Ile Asp Ser
165 170 175
Asn Asn Phe Asn Glu Ser Tyr Ile Tyr Asn Phe Ile Cys Ser Glu Ser
180 185 190
Gly Lys Ile Arg Glu Ser Lys Asn Tyr Lys Ile Arg Thr Asn Arg Asn
195 200 205
Arg Ser Asn Leu Ile Ser Ser Lys Asp Phe Asp Ile Thr Gln Asn Tyr
210 215 220
Asn Gln Leu Trp Ile Gln Cys Asp Asn Cys Tyr Gly Leu Met Tyr Lys
225 230 235 240
Lys Val Lys Met Asn Val Cys Glu Gln Cys Gly His Tyr Leu Lys Met
245 250 255
Ser Ser Ser Glu Arg Ile Glu Leu Ser Ile Asp Pro Gly Thr Trp Asn
260 265 270
Pro Met Asp Glu Asp Met Val Ser Ala Asp Pro Ile Lys Phe His Ser
275 280 285
Lys Glu Glu Pro Tyr Lys Asn Arg Ile Asp Ser Ala Gln Lys Thr Thr
290 295 300
Gly Leu Thr Asp Ala Val Gln Thr Gly Thr Gly Gln Leu Asn Gly Ile
305 310 315 320
Pro Val Ala Leu Gly Val Met Asp Phe Arg Phe Met Gly Gly Ser Met
325 330 335
Gly Ser Val Val Gly Glu Lys Ile Thr Arg Leu Ile Glu Tyr Ala Thr
340 345 350
Asn Gln Cys Leu Pro Leu Ile Leu Val Cys Ser Ser Gly Gly Ala Arg
355 360 365
Met Gln Glu Gly Ser Leu Ser Leu Met Gln Met Ala Lys Ile Ser Ser
370 375 380
Val Leu Cys Asp Tyr Gln Ser Ser Lys Lys Leu Phe Tyr Ile Ser Ile
385 390 395 400
Leu Thr Ser Pro Thr Thr Gly Gly Val Thr Ala Ser Phe Gly Met Leu
405 410 415
Gly Asp Ile Ile Ile Ala Glu Pro Tyr Ala Tyr Ile Ala Phe Ala Gly
420 425 430
Lys Arg Val Ile Glu Gln Thr Leu Lys Lys Ala Val Pro Glu Gly Ser
435 440 445
Gln Ala Ala Glu Ser Leu Leu Arg Lys Gly Leu Leu Asp Ala Ile Val
450 455 460
Pro Arg Asn Leu Leu Lys Gly Val Leu Ser Glu Leu Phe Gln Leu His
465 470 475 480
Ala Phe Phe Pro Leu Asn Thr Asn
485
<210> 5
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
ctagtctaga atggaaaaat cgtggttcaa t 31
<210> 6
<211> 41
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
aaactgcaga tttgtgttca aaggaaaaaa agcatggagc t 41
<210> 7
<211> 159
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
atggcccttc aagctgcttc tttggtctcc tctgctttct ctgtccgcaa agatggaaaa 60
ttaaatgctt cagcatcatc atcattcaaa gagtctagtc tgttcggtgt ttcactttcg 120
gagcaaagca aagctgactt tgtctcttcc tcattgaga 159
<210> 8
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
tacgagctca tggcccttca agctgcttct 30
<210> 9
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
ctagtctaga tctcaatgag gaagagacaa ag 32
Claims (3)
1.蛋白质在提高高温下转基因植物育性中的应用,其特征在于,所述蛋白质的氨基酸序列如SEQ ID NO.3所示;所述植物为拟南芥。
2.编码如权利要求1中所述的蛋白质的核酸分子在提高高温下转基因植物育性中的应用,其特征在于,所述核酸分子的核苷酸序列如SEQ ID NO.1所示;所述植物为拟南芥。
3.与权利要求2中所述的核酸分子相关的生物材料在提高高温下转基因植物育性中的应用,其特征在于,所述生物材料为下述1)至7)中的任一种:
1)含有如权利要求2所述核酸分子的表达盒;
2)含有如权利要求2所述核酸分子的重组载体;
3)含有1)所述表达盒的重组载体;
4)含有如权利要求2所述核酸分子的重组微生物;
5)含有1)所述表达盒的重组微生物;
6)含有2)所述重组载体的重组微生物;
7)含有3)所述重组载体的重组微生物;所述植物为拟南芥。
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Baoye He et al.Effects of inefficient transcription termination of rbcL on the expression of accD in plastids of Arabidopsis thaliana.Photosynth Res.2015,摘要,第4页左栏第1段-右栏最后1段,第5页左栏第3段. * |
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Maria N. Danilova et al.Differential impact of heat stress on the expression of chloroplast-encoded genes.Plant Physiology and Biochemistry.2018,第129卷摘要,第94页右栏第2段,第96页右栏第1-2段,图2B. * |
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