CN109207634A - 海州常山干旱胁迫下荧光定量内参基因及其专用引物和应用 - Google Patents

海州常山干旱胁迫下荧光定量内参基因及其专用引物和应用 Download PDF

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CN109207634A
CN109207634A CN201811441119.7A CN201811441119A CN109207634A CN 109207634 A CN109207634 A CN 109207634A CN 201811441119 A CN201811441119 A CN 201811441119A CN 109207634 A CN109207634 A CN 109207634A
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clerodendron trichotomum
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岳远征
杜菊花
王良桂
杨秀莲
施婷婷
华雅洁
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Nanjing Forestry University
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Abstract

本发明公开了海州常山干旱胁迫下荧光定量内参基因及其专用引物和应用，该荧光定量内参基因为Actin、AP‑2和RAN基因，其中，Actin基因的基因序列如SEQ ID NO.1所示，AP‑2基因的基因序列如SEQ ID NO.13所示，RAN基因的基因序列如SEQ ID NO.6所示。本发明通过从转录组数据以及前人研究的内参基因中筛选出表达较为稳定的候选基因，通过软件对其稳定性进行评估，公开了3个适合于海州常山干旱胁迫下表达最稳定的内参基因，并用这些基因为基础设计了实时荧光定量PCR引物，填补了海州常山干旱胁迫下没有内参基因的现状，为海州常山干旱胁迫下相关功能基因的精确定量提供有力支持，可提高研究的稳定性、重复性和可靠性。

Description

海州常山干旱胁迫下荧光定量内参基因及其专用引物和应用
技术领域
本发明属于植物分子生物学领域，具体涉及海州常山干旱胁迫下荧光定量内参基因及其引物和应用。
技术背景
海州常山(Clerodendrum trichotomum Thunb.)又名臭梧桐，马鞭草科(Verbenaceae)大青属(Clerodendrum L.)落叶灌木或小乔木，较耐旱，耐盐碱性较强，有一定耐寒性，极少病虫害，适应性强。此外花形奇特美丽，花期长，果实色彩鲜艳且秋冬经久不落，是优良的秋季观花、观果树种。现有的研究结果表明，海州常山不仅具有非常优良的耐盐碱能力，还有较强的耐早性，是一种对不良环境具有较强抗性的优良园林植物。现在关于海州常山干旱胁迫下基因的研究还未曾有报道。探究海州常山干旱胁迫情况下基因表达的差异的研究过程中，涉及到运用实时荧光定量技术分析和验证基因的表达水平就需要稳定可靠的内参基因，因此筛选海州常山干旱胁迫条件下稳定表达的内参基因是实时荧光定量结果准确的关键因素。
发明内容
发明目的：针对现有技术中存在的不足，本发明目的在提供适于海州常山干旱胁迫条件下的内参基因，该基因能够满足实时荧光定量检测海州常山转录表达水平的要求，提高了海州常山基因表达分析研究的稳定性、可靠性和效率。本发明的另一目的是提供一种上述内参基因的专用引物。本发明还有一目的是提供一种上述内参基因或专用引物的应用。
技术方案：为了实现上述发明目的，本发明采用的技术方案为：
海州常山干旱胁迫下荧光定量内参基因，为Actin、AP-2和RAN基因，其中，Actin基因的基因序列如SEQ ID NO.1所示，AP-2基因的基因序列如SEQ ID NO.13所示，RAN基因的基因序列如SEQ ID NO.6所示。
所述的海州常山干旱胁迫下荧光定量内参基因的专用引物，Actin基因的引物序列如下：
Actin正向引物5’-CGATTCTTGCTTCCCTCAGTA-3’
Actin反向引物5’-ACCATTTTCGCCGCCTTA-3’。
所述的海州常山干旱胁迫下荧光定量内参基因的专用引物，AP-2基因的引物序列如下：
AP-2正向引物5’-CCACAACTAACGCCTCCACC-3’
AP-2反向引物5’-AAATTCACCCTCCGACTCCC-3’。
所述的海州常山干旱胁迫下荧光定量内参基因的专用引物，RAN基因的引物序列如下：
RAN正向引物5’-GGAGACGACAGTTTGGGTGC-3’
RAN反向引物5’-TGCCATACTTGCGGGTGAA-3’。
所述的海州常山干旱胁迫下荧光定量内参基因在海州常山荧光定量中的应用。
所述的Actin基因的引物序列在海州常山荧光定量中的应用。
所述的AP-2基因的引物序列在海州常山荧光定量中的应用。
所述的RAN基因的引物序列在海州常山荧光定量中的应用。
本发明通过GeNorm、NormFinder和BestKeeper这三个软件对17个内参进行基因稳定性评价，综合软件分析结果，筛选出在干旱胁迫条件下稳定表达的内参基因。共获得三个稳定表达内参基因分别为Actin、AP-2和RAN。
有益效果：与现有技术相比，本发明的优点在于：本发明通过海州常山的转录组测序库，并参考了常用的内参基因，设计出了适用于海州常山干旱胁迫下的内参基因，同时揭露了这些内参基因序列，并依此设计了实时荧光定量引物，不仅解决了现有海州常山检测中没有内参基因的现状，而且所设计的实时荧光定量引物用于海州常山基因表达分析时引物特异性强，从而能够大大的提高了采用实时荧光定量检测海州常山基因时的检测效率，并提高了检测结果的可信度。
附图说明
图1是琼脂糖凝胶电泳检测RNA质量胶图；图中，M为DNA2000Marker，用到1-9均为样品的RNA；
图2是17个候选内参基因的Ct值；
图3是GeNorm确定用于准确定量分析最优的内参基因数目；
图4是GeNorm软件对17个候选内参基因表达稳定值(M)排序，稳定值越低，基因表示越稳定结果图；
图5是应用ClNHX1验证所筛选出的内参基因结果图。
具体实施方式
下面结合具体实施例对本发明做进一步的说明。
以下实施例所使用的主要供试材料为：采用种植于南京林业大学白马基地的泰安种源、盐城种源。采集组织材料植株插穗进行扦插，基质使用珍珠岩∶蛭石∶泥炭∶沙子＝1∶1∶1∶2，栽于培养室(25℃，60％RH)，定期浇灌养护，保证正常生长。胁迫处理均在人工气候培养箱(宁波东南仪器有限公司)中完成，昼夜温度及时间设定为25℃/21℃和13h/11h，湿度为60％RH，光强为100001ux。待需要进行干旱胁迫处理时，将海州常山扦插苗移栽于不含糖、琼脂、有机物及碳源的1/4MS培养基，每瓶加入150mL的培养基，每5天更换一次培养基，适应一个月。进行干旱胁迫时，在培养基中加入15％PEG，并于0h、2h、6h、12h、24h、48h、72h采集海州常山的嫩叶。叶片均采自于从上至下数2-4对叶片，每次采样均有三个生物学重复，样品用液氮速冻，在-80℃下保存。
实施例1
1、植物组织总RNA提取及cDNA的合成
用EASYspin Plus植物RNA提取试剂盒(艾德莱生物技术有限公司)提取总RNA，用分光光度计(梅特勒-托利多集团)检测RNA浓度和质量，利用1.5％的琼脂糖凝胶电泳检测总的完整性。利用北京全式金反转录试剂盒(全式金生物技术有限公司)，按说明书进行RNA的纯化和合成cDNA第一条链。
2、内参基因的选择及引物的设计
根据文献查询其他植物中主要的内参基因并最终设计出本实施例所需的17个内参基因，分别为Actin(SEQ ID NO.1)、PP2A(SEQ ID NO.2)、RPL(SEQ ID NO.3)、PK(SEQ IDNO.4)、18S(SEQ ID NO.5)、RAN(SEQ ID NO.6)、APT(SEQ ID NO.7)、SAND(SEQ ID NO.8)、PROF(SEQ ID NO.9)、MDH(SEQ ID NO.10)、EF-1A(SEQ ID NO.11)、UBC-E2(SEQ ID NO.12)、AP-2(SEQ ID NO.13)、HSP70(SEQ ID NO.14)、TUA(SEQ ID NO.15)、UBQ(SEQ ID NO.16)、H3(SEQ ID NO.17)。首先在本申请人已经获得的海州常山转录组数据库中寻找各候选基因的保守序列，根据基因保守序列用Primer Premier 5.0设计引物，再用Oligo7进行分析。引物(表1)均在南京金斯瑞公司合成，通过普通PCR初步筛选引物，利用琼脂糖胶和凝胶成像系统观察PCR产物的条带，检测产物特异性。选择条带大小正确，条带特异性好，没有引物二聚体的引物，近一步通过RT-qPCR进一步检测引物特异性，选择峰图单一，没有杂峰，阴性对照无峰的引物作为最终的引物。
表1内参基因及其对应的引物
3、内参基因引物标准曲线的建立
对每一对内参基因的引物制作各自的标准曲线，计算对应引物的扩增效率。将反转录的cDNA混合后，以5为倍数稀释成5个梯度(1、1/5、1/25、1/125、1/625)，作为建立标准曲线的模板。同时，以dd H2O作为阴性对照模板，来检测实验过程中的试剂或者人为污染。所有的样品均做3次重复，以确保实验数据的可信度。用Applied Biosystems StepOne(Thermo Fisher Scientific，USA)进行RT-q PCR，利用得到的数据结果求得各候选基因的扩增效率及斜率，荧光定量PCR扩增效率要求所选引物的扩增效率在90％-110％之间。
4、荧光实时定量
使用Applied Biosystems StepOne(Thermo Fisher Scientific，USA)进行RT-qPCR，以叶片样品第一链为模板进行定量分析，首先将模板稀释10倍。利用Premix ExTaqTM(TakaRa，Japan)进行实时荧光定量，反应体系为总体积为10uL，正反向引物0.4uL、SYBR5uL、cDNA1 uL、校正液0.2uL、超纯水3uL。扩增程序为95℃预变性30s，1个循环，95℃变性5s，60℃退火30s，共40个循环，再运行溶解曲线阶段的1个循环，95℃15s，60℃1min，95℃15s，每个样品设3个重复，并设阴性对照。
5、数据处理
内参基因稳定性分析，利用软件GeNorm，NormFinder，BestKeeper和在线分析工具RefFinder(http：//150.216.56.64/referencegene.php)综合分析内参基因的表达稳定性。筛选出稳定的内参基因。
6、结果
1)RNA提取质量及引物特异性检测
各样品的OD260/280、OD260/230均符合要求，琼脂糖凝胶电泳图检测显示28S及18S条带清晰，其灰度比约为2∶1，5S条带亮度较暗，无降解现象，满足要求(图1)。17对引物在荧光实时定量中熔解曲线均只有明显的单一峰，且电泳检测也只有单一条带图，说明所用引物可特异扩增各内参基因的相应产物，无引物二聚体，且每个待测样品扩增曲线的重复性好，模板能特异性扩增，实时荧光定量结果准确可信。
2)内参基因Ct值分析
Ct值与基因的表达量呈反比，Ct值越大，基因的表达量越低，反之，Ct值越小，代表基因的表达量越高。17个内参基因Ct值的平均值在18.70-26.47之间，其中Actin表达量最高，APT和PROF的表达量较低(图2)。
3)软件分析
GeNorm软件分析：GeNorm软件是根据平均变异度M值来衡量基因的稳定性，软件默认的取舍值为1.5，高于1.5的基因不适合作为内参使用，M值越低表示基因越稳定。此外GeNorm软件还根据候选内参基因的配对差异值Vn/Vn+1来确定合适的内参基因个数，当Vn/Vn+1＜1.5时，采用n个内参基因个数。根据软件，计算出V3/V4＝0.136＜1.5(图3)，所以适用于海州常山干旱胁迫下应选用三个内参基因，分别为Actin、UBC-E2和PP2A(图4)。
NormFinder软件分析：NormFinder软件是根据计算候选内参基因的稳定值进行评价，稳定值越高稳定性越差，具有最小稳定值的基因为最稳定的基因。结果如表2所示，AP-2的值为0.283，为最稳定的基因，H3稳定值为2.427，为稳定性最差的基因。
表2 NormFinder软件分析结果
排名 1 2 3 4 5 6 7 8 9
基因 AP-2 RAN Actin SAND UBC-E2 PP2A UBQ HSP70 RPL
稳定值 0.283 0.42 0.427 0.496 0.601 0.634 0.716 0.742 0.796
排名 10 11 12 13 14 15 16 17
基因 EF-1A TUA MDH PK 18S APT PFOF H3
稳定值 0.83 0.833 0.841 0.896 0.924 1.511 1.594 2.427
BestKeeper软件分析：BestKeeper软件是基于内参基因Ct值的标准差(SD)和调节系数标准差SD(±x-fold)来评判基因的表达稳定性，直接对基因表达Ct值进行分析。SD值越小，表达越稳定，程序默认临界值为1，当SD值大于1时，认为该基因表达不稳定。分析结果如表3所示，显示从PROF、H3和APT，SD数值大于1，其余基因SD数值均小于1，基因的稳定性为AP-2＞Actin＞MDH＞RAN＞UBC-E2＞UBQ＞RPL＞SAND＞PP2A＞EF-1A＞TUA＞HSP70＞18S＞PK＞PFOF＞H3＞APT，表达最稳定的是AP-2，最不稳定的为APT。
表3 BestKeeper软件分析结果
排名 1 2 3 4 5 6 7 8 9
基因 AP-2 Actin MDH RAAN UBC-E2 UBQ RPL SAND PP2A
SD 0.4 0.55 0.57 0.75 0.76 0.79 0.8 0.81 0.83
CV 1.73 2.95 2.41 3.06 3.54 3.24 3.65 3.12 3.7
排名 10 11 12 13 14 15 16 17
基因 EF-1A TUA HSP70 18S PK PFOF H3 APT
SD 0.84 0.88 0.89 0.95 0.97 1.15 1.56 1.58
CV 3.87 3.79 4.29 3.81 4.28 4.03 7.06 5.97
4)内参基因稳定性验证
综合考虑Ct值和BestKeeper、GeNorm和Normfinder 3个软件的分析结果得出，Actin、AP-2和RAN是适合于海州常山干旱胁迫条件下适用的内参基因(表4)。
表4综合排名
选用ClNHX1(SEQ ID NO.18)，来验证所筛选的内参基因，荧光定量PCR程序上同，并用表达不稳定的H3基因来做比较，发现用所筛选出的内参基因来作为内参时ClNHX1有着相似的变化趋势，但用H3基因来做内参时，ClNHX1基因表达情况不同，说明筛选出的内参基因是准确可靠的(图5)。
序列表
<110> 南京林业大学
<120> 海州常山干旱胁迫下荧光定量内参基因及其专用引物和应用
<130> 100
<160> 52
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1283
<212> DNA
<213> 海州常山的Actin基因(Clerodendrum trichotomum Thunb)
<400> 1
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aacgttcagc gcagtccaaa atactacagc gttaaggaca cgtctagaaa accataaacg 120
attttttttt ttagaaaagc aaatataaat agtacctagg cataccaacc aaaacaaaag 180
accatataaa ataacttcat gcactcaaaa ctgcaatcga actcacacgg actccaatag 240
aagcatgaat aaatgatacc attttcgccg ccttagaagc actttctgtg aacaatagca 300
ggacctgatt catcgtactc tccttttgca atccacatct gttggaaagt actgagggaa 360
gcaagaatcg accctccaat ccagacacta tacttccttt caggtggggc aaccaccttt 420
atcttcatac tactcggagc aagagccgta atttccttac tcatacgatc agcaataccc 480
gggaacatag ttgaaccacc actgagcaca atgttcccat acagatcctt cctaatatcc 540
acatcacatt tcattataga attgtaagtt gtttcatgaa ttccggcagc ttccattcca 600
atcagtgatg gctggaacaa cacttccgga caacggaacc tctcagcgcc gatggtgatg 660
acctgtccat ccggaagctc gtagctcttc tccaccgcag agctgttctt cgccgtctct 720
gtttcctgct cgaaatcaag tgccacataa gcgagcttct cctttacatc gcggacaatt 780
tcccgctcag cggttgttgt aaacatgtaa cctctttcag tcagaatctt catgaggtaa 840
tcggtgaggt cacgaccagc gagatccaat ctcagaatag cgtggggaag ggcgtaccct 900
tcataaatag ggacagtatg gctcacaccg tcaccagaat caagcactat acctgttgta 960
cgaccactag cataaagaga gaggacagcc tggatggcaa catacatggc cggcacattg 1020
aaggtctcga acatgatttg agtcattttt tctctgttcg ctttagggtt gagaggggcc 1080
tctgtgagga ggacagggtg ctcttcagga gcgacacgca gctcattgta aaatgtatgg 1140
tgccaaatct tctccatatc atcccagttg ctcactattc catgctcaat tgggtacttc 1200
aatgtgagga tacctctttt ggattgggct tcatcaccta cataagcatc tttttggccc 1260
attccaacca tgacacctgt atg 1283
<210> 2
<211> 1437
<212> DNA
<213> 海州常山的PP2A基因(Clerodendrum trichotomum Thunb)
<400> 2
attcccagaa actgtcatgc aagagattgt aaaaatggtg tccataaatt tgtttcgtac 60
actctcgtcg cagcctcgtg aaaacaaagt cttggaagct tttgatgtgg atgaagagga 120
acccttgatg gatcctgcct ggccccattt gcaagttgtg tatgagtttc ttctaaggtt 180
tgtggcttca ccagagacag atgcaaagtt agcaaaacgt tacattgatc actcctttgt 240
tttgaggtta ttagatcttt tcgactctga agatcccaga gaaagggact acctaaagac 300
tgtccttcac cgcatatatg gaaagttcat ggttcatcgt cccttcatca ggaaatcgat 360
caataatata ttctaccact tcatatttga aacagagaag cataatggga tcgctgaact 420
gttagaaatt ctgggtagca ttataaatgg ttttgcattg cctttgaaag aagaacataa 480
acttttcctt gttcgggcac tcattccgct tcacaagcca aagtgcatcc ctatgtacca 540
tcaacaacta tcgtattgca taacacaatt tgtggagaag gactgcaaac ttgctgatac 600
tgtcataaga ggcttactca ggtattggcc gatcacaaac agttcaaagg aggtaatgtt 660
cttgggtgag ctagaggaag tgttggaagc aactcagcct ccagagtttc agcgttgtat 720
ggttcctttg ttccgccaga ttgctcgttg cctgagcagt tcgcattttc aggttgcaga 780
aagggctctc tttttatgga acaacgatca catcgagaac ctgatcaaac aaaaccgtag 840
agtgatccta ccggttatac tccctgcatt ggagaagaac tcaagaaccc attggaacca 900
ggcggtacag agtttgacca tgaatgtccg taaaatcttc tccgatgctg atcctgagct 960
ctttgaggaa tgtttgatca aattccaaga agacgaagca agggaagaag aaaccaaaaa 1020
gaaacgggaa gcaacatgga aacgtctaga agaaatagcc gcaatgaaag cttctagtaa 1080
tgaaccggtc cttgtttctc ccaggacaag caccaagacg ccctccggct aaacatggaa 1140
ggcttcgttt gagtcgcttg aattgtggaa ttatgtattt gcccctactc atgcttctgg 1200
ttttttttgt ccggttgggt cgagcagggt gaaggaaact aatatctttt tgggggttat 1260
gatgaagcgt tgtttttgat tgtaagaaat agtgggtgta catatagaat tgtgggggcc 1320
agatactgat gcggttttag aagtgctgat gttctatggg ggaatatttg ctggcaaatt 1380
tgccctactt tcatctttag ctctttagaa tgatgttaaa attttaagtt atatttt 1437
<210> 3
<211> 1554
<212> DNA
<213> 海州常山的RPL基因(Clerodendrum trichotomum Thunb)
<400> 3
ttaaagcgaa gtctcactag atctgggtcg ttgtctaact gagtgataat cttatataat 60
taaccaaagc ctgcatatag aatagggttt tgagcctctg gaattttact ttgcggcggt 120
cagagtaatt aggcccaaga aactatgtct cacagaaagt ttgagcaccc aagacacgga 180
tctttggggt ttctaccaag aaagagggcc tctcgtcaca gaggaaaagt gaaggccttt 240
cccaaggatg atccaacaaa accttgcagg ttgactgctt ttcttggata caaagcaggg 300
atgacacata ttgttcgtga ggttgaaaaa cctgggtcca aactccacaa gaaggagaca 360
tgtgaagctg taactattgt tgaaacccca ccaatgattg ttgttggtgt tgttgggtat 420
gtcaaaacac cacgaggcct ccgttgcttg aatactgttt gggctcaaca tcttagtgaa 480
gacctcaaga ggaggttcta caaaaactgg tgcaagtcca aaaagaaggc tttctcaaag 540
tattcaaaga agttcgacag tgaagaagga aagaaggata tccaagccca gttggagaaa 600
atgaaaaagt actgcactgt tgttcgtgtt cttgctcata cccagattcg aaagatgaaa 660
gggcttaagc agaagaaagc ccacttgatg gagatccaag tcaatggtgg cgatgtagct 720
aagaaggttg actatgcata cagctttttc gaaaaacagg tccccattga tgctgttttt 780
gggaaagatg agatgatcga cattatcgga gtgaccaagg ggaagggtta cgagggcgtg 840
gtgacccgtt ggggtgtgac tcgactccca cgcaagaccc accgtgggct tcgcaaggtt 900
gcctgtattg gcgcatggca tcctgctagg gtttcattca ctgttgctag ggctggacaa 960
aatggttacc atcacagaac tgagttgaac aagaagatct acaaattggg caaggctggt 1020
caagagtctc actctgccat gacagagttt gataggaccg ataaagatat tacaccaatg 1080
ggagggttcc cacactatgg tgtggtgaag gatgactatc tgttgataaa gggctgctgt 1140
gtgggaccca agaagcgcgt cgtcacattg aggcagtctt tgttgaacca gacgtctagg 1200
gttgcgttgg aggagatcaa actcaagttc attgatacgt cttctaaatt cggccatggc 1260
cgcttccaga ctatggacga gaaagctaag ttttttggca agctcaaggc ttaaaatgac 1320
gtcgattggg tggggatata atggaatcgg tttttatcgt tcattttcct ttataacttt 1380
gttggtttgc cttttttccc ttgtaagctg caggtcttga aatcgtccaa agaactttaa 1440
atagattttg catttgtgtt aaacctgtac cctgcacaat tacaatcatc tacatgttct 1500
catttgcgtt cataagatgc atgtactggt aaatggctat acttaattaa gatg 1554
<210> 4
<211> 1758
<212> DNA
<213> 海州常山的PK基因(Clerodendrum trichotomum Thunb)
<400> 4
agtgataaca gtaatctcct aaagtctcaa tcccatacca caagtaccca ccacttccaa 60
tctctctcct ccaattacac tctctcttcc acacatactc acgcactttt ctgagcgagg 120
cgcccacaca ttgaacacag tgagaatggc gtccacagct gcatcatcag ccttctgcgg 180
catccgcaag gcatcgaccg ccgctgccgc caaggcttct ctcaatgccc ctaccaccta 240
cttcctcccc aaaacccccc tccgccgcct cggattcgcc gccggagccg ccgaaccgat 300
cctatctcac catgtcgcca ctaagatgcg atcatttggg aattccggca aaccaattag 360
aggcgtggcc tccatggcga agaagagcat cggagacctt ggtgctgctg acctcaaggg 420
caagaaagtc tttgttaggg ctgacctcaa tgtgcctctt gatgacaacc agaatattac 480
tgatgatact aggattagag ctgctgtccc gacaattaag catctgatta acaatggtgc 540
taaagtcatt ctttctagtc atttgggacg accgaaaggc gtcacgccaa aatacagtct 600
ttcacctctt gttcccaggc tatctgaact tctgggcatt caggttgtga aggccgatga 660
ctgcattggt ccagaggttg aaaaattggt cgcttcactg cctgaaggtg gtgtactcct 720
gctcgagaat gtaaggtttt ataaagagga agagaagaac gagcctgagt ttgcaaagaa 780
acttgcctcg ttggctgatc tttacgttaa cgatgcattt ggtaccgcac acagagcaca 840
tgcctcgact gagggagtca caaagttctt gaagccctct gttgctggtt tcctattaca 900
aaaggagctt gactatcttg ttggggcagt ttcgaaccca aaaagaccat ttgctgccat 960
tgttggtggt tctaaggtct cctccaagat cggagtaatt gaatcacttc tagagaagtg 1020
tgatattttg ctattgggag gaggaatgat atttacattt tacaaagcac aaggactttc 1080
agtggggtcg tctctagtag aagaagacaa gctaaatcta gcaacatcgc tccttgagaa 1140
ggcaaaggcc aagggggtga gcctcttgct gcccactgat ttggttatag ctgacaagtt 1200
cgctcctgat gctaatagca agattgtggc agcatctgct attccagaag gatggatggg 1260
attggatatt ggcccggatt ctataaagac attcagtgat gcactgggga ctacacaaac 1320
tgttatttgg aatggaccga tgggagtgtt tgaatttgac aagtttgctg ttggcacaga 1380
ggcgattgca aacaagctag cagaacttag tgggaaggga gtgacaacaa ttattggtgg 1440
aggagattca gtagcggcag tggagaaagt tggagttgct aatgtgatga gccacatatc 1500
gactggtggt ggtgccagtt tggagctctt ggaaggcaaa gaactgccag gtgttcttgc 1560
ccttgatgaa gccacgcctg ttgccgtgta gtgtgtaaaa aacaccattc ttctcgatta 1620
ttttaacacc catctctaat aatgtctcat tactagcttg taaacgagaa atttcttggt 1680
atacgtggtc atgctataga tgttttacct tgcctccttt tcttgtcatg ttaatatcac 1740
accatggaat ggatccgt 1758
<210> 5
<211> 1308
<212> DNA
<213> 海州常山的18S基因(Clerodendrum trichotomum Thunb)
<400> 5
cgaatgctgt taaatttgaa tggtttaaat aaaaacatat taagagagat tacgcaagct 60
taaatctctg gcaacaatca actttcctcg aataattgta aagggtgaag aagaaaaaaa 120
aatcaaaaag caaaggccaa aatttggtat gaatagcaag tagcaacaca agcaaatttg 180
cttgtgtgga cctaacatct tctgtgccac aatttccata attggatcgc ttgcgcgact 240
atagagattc ttattagaac cgagcttttc ttttccgagc agtgtatttt gaatctggtg 300
gcactatatt cccctttctg cgcatttgtt ccttcttacg cagaacccat tctttcccct 360
tcaccttctt gtttagtttt tgccttttcc ttgctctctg ccggtctgat acaaaaactg 420
tctgttcttc atcatcactg ctatcttcat cagaactctc tccatcttca cctttcgcct 480
ttggagtcgc actactcaga gatggtgggc cacaagtgag cacgaggtat tctttcctcg 540
atttcgtact gttaccaaga agaagtatta cagttaacgt gacaacttat cagaactcca 600
gtgcagggaa gttgattcaa acctccaagg aatttacaag aataaaaccc tatgcggata 660
atcaactact acaccaccag aaaacccagc acgcatagca aatcccaaga tcagctcacg 720
ttgagccaaa ttttcaggat atacctgtag cactgcccta gctcctcgtg ctaaacatct 780
gtataatgac ccaaagaagg ccttcagtct caagcgtggc tcatgagaag atttgtcagc 840
attgcataac cactgaacag ctgagatact tattgcgcca tcaataactc cagggcgaag 900
ccctaagccc tgacccatgt caccaagtat aaggtcaccc tcagcttcac gctccaatgc 960
aatatctagc attgatcctg atatatctaa accgatccat tggtgcccat ttccagccaa 1020
tgtctcccca ctaagtccag atccacaacc gatatcgaga agtaatcttg gaacgccatc 1080
atcaggcaaa gcgagaagct ccaatgccct ctcggaaagt ttagcctgaa tttcgatgat 1140
acgagaagaa gaggtgtact tacgagcttc ctcgtcgtta taaaatatct caggcggtgc 1200
ttgtagctct ggtcgagagg ccattgtttc tttccttgtt cttcgaaaca gtactcagag 1260
ggtggctgaa ggctgctttg ttcggtgatt gtccggtttt gtctgatg 1308
<210> 6
<211> 2008
<212> DNA
<213> 海州常山的RAN基因(Clerodendrum trichotomum Thunb)
<400> 6
ccccaacatg tgggacccat aaattagaat atgcatacga agtaacagtg aattaccaaa 60
agtaccctca gctgatacat aatgcaaaat aatcaaaagt agagagatca agggcaaaaa 120
tgtcacataa aatcaaacac agcacacttt tggcaagcag ttggcttcag gcttaaacag 180
cgccatctcc tgctcaaatc tagcagaccc accaaagaca tcaaactaaa acctaatgat 240
ctaaatgaca aaactatctt aatctttgtc ttcttcattg acatcaagat tcttgagctt 300
cttttccaat tcatcctgac taccttcgtc ttcatccccc ttgtcgtcaa aatcgtttcc 360
ttcagggtcg ttctcatcca acggcccaag tttctcagga caatctttaa agattatcct 420
caactcatcg agatcttcct cagaaatgaa gttcccgttt acatccaata gcttgaaacc 480
aggcaaacga accaaagcca gagccaactt cttaaccgag gtagttagtg agttctggct 540
catatcaact tccctcaatt gctcttgccc atcttcaaat gctttgctta tctgaatcgc 600
accttcatcc ttaaggtcat tctctgagag gttcaacttg acaatagttt tctttctaga 660
gatgcaggcc gccaagctag gagcagcttc agcagttata tcatttccag ccatctccaa 720
tactgcaagt gatggtgctg actccttgag agcgtcggct attgctattg ctccatcatc 780
ttcaagattc gagtagctta aataaatctc agttagattc ccatgtttgg agagagcgtc 840
gcttagcttc aaaccagcct ccactccaaa catattgtcc cgaagatcga ttttcttcag 900
attcttacat aattccaggg cttcagtcaa cgcaacgcct ccttcagaac caatacgggt 960
agatgagcat cggaaatcct ccaataaagg gcaacgtttc agtatctcgg aaatggcaat 1020
agcgccttca tctcctgtca tattgttatg aaaatggagg attctaagcg aatctgtaga 1080
aggaaccaac tcacaaacag cttgtgcggc ttccttcgat atcccatcat tcatcagata 1140
aagttcctct aagctagtct gagatttcaa gagcttcccg aatgctctga ctcccttctc 1200
acctaaggca ttatcggaga tattgagata tttcaaatga gaaccttcta gagcttcaga 1260
gaagatcttc atgacatcaa gagcttctgc ttctggtcgc cccgcaacaa aatcagacag 1320
atcgacttcc ttcagttggt tcttgattga cgccaagata ggtccagcaa cattggctgc 1380
cccaagacca aagcttcgat tgctaaagca aatcttggtg taagaattcc caggttcttt 1440
caaggggctt aaaagtgttt gagcctcatc tgcttcaata aatgctcgct gacccttgga 1500
aatatcgaaa aatgtctcct gcaaaaccga attgacctca gatttcagta cctgcttttc 1560
ctctgtccca gagcctccct tgagaacatc cagtattagc ttgctgcatt ccttggcata 1620
ccattgtaca gcagaactgc catcaccatc gggttcttgt tcataatgct gatttgccga 1680
agaaaatgct gattcttcaa tttgtttggc atgttttgaa gcctcagctt gactaagagt 1740
gccatacttg cgggtgaaaa ctgttggcgt cgagagattg tttgtcattc gctctacaag 1800
catcaacctc gtgttctggc tagggggcca cagtttaatg gttatgtgcc ggcgttctcc 1860
atttggtgtt ttagcatcca tcgcacccaa actgtcgtct ccttcctgac gaatttttca 1920
atttgcagct cttcggactt cgactgtgta ctgcctgtgt atgtataggc aagaggggag 1980
aagtgaagga aaactaaaat gaaatgag 2008
<210> 7
<211> 1149
<212> DNA
<213> 海州常山的APT基因(Clerodendrum trichotomum Thunb)
<400> 7
aaaatctaga gatttaggca aatccattag aaaacaagca tcctgtcttc aattgattct 60
aaaaatccca ctaatgtacg ctgtatctgt atctaagcca tctctctccc tatcaagtga 120
tgtcattgat taaacaaatc ttgagtttta ggcaaatcca ttagaaaaca agcaccctca 180
tggacttcaa aaatcccact aatgtatgct gcatctgtat ctaagctatc cctcaaactc 240
ctcaaggcta gagttgtctc accaaagtac ttgcgcagtg ttaatttctg cttacctcac 300
accgtagcga acactccgct gacctacgtc ggatcaattg gtggcgcagc cgtaccgaaa 360
cgtcggccgg tgttcgtttc aaacatggca tcctcaaatg ctgaaagcat tgatgagcgc 420
atccggcgga ttgcctccac gattcgggtc atacccgact ttccaaaacc tgggatcatg 480
tttcaagata taacgacgtt gttgcttgat cctaaggcgt ttaaggatac gattgatttg 540
tttgttgaga gatataaatg ccaaaacatc aatgttattg ctggtgttga ggcaaggggt 600
tttatattcg gtcctcctat tgcattggct attggtgcaa aatttgttcc tatgaggaag 660
cccaaaaagt tacctgggga agtaatatca gaagagtact ctttggagta tggaactgac 720
ataatggaga tgcatgttgg agccgtacaa cctggggaac gtgcagttgt tgtagatgat 780
cttatcgcaa cagggggtac cttgaatgct gctattcgcc tacttgaacg cgttggagtc 840
catgtggttg agtgtgcttg tgttattgaa ctggagggac tgaagggtcg ggacaagtta 900
ggggagaatc cactgtttgt tcttgtgagc tcaccttgaa gaaaagagta tctacaacaa 960
caataaattg ctgttaacat ctgtatgagt tgatagcatg tgcttgttcc tttctatggg 1020
atatacacat ttcaaatgga tgatggtgtg ttgtatgttg agtgaaattg cttttctttt 1080
ttttttggat gagtcatgtt caggccaccc tctcctgctt gcggattgtt actcaacctg 1140
tcaaataag 1149
<210> 8
<211> 2473
<212> DNA
<213> 海州常山的SAND基因(Clerodendrum trichotomum Thunb)
<400> 8
gtaatttgtt tctgtctccg ccatccgtga ctggcagatg cgagtgcgac accatcactg 60
aacttcaaat gttttccttt aaactctaat ttcttagcga ttatccaatc caattctcgt 120
tcgggattgt agattgattg acaatgtctt ccaattcaaa tatttcatct gcgagttcct 180
ctactaatgg agatgacggg aaccctaatt cgtatacaaa aaaggagccg gagaatctgc 240
cggaacccgc actcaacacg agtccggact tgatcgacca gtcgcttgat gcccttgaga 300
atcaattggc ctcaatatca atgtcctccc gttccccgac ggcgactctt gacgatgaag 360
agttcgtcga agaagaagag cagtacctgg agcatccaac aatcataaat ggatcgttga 420
gtgaagaaaa tggagccgga ggatcaagtg atgagcatat tgagtgttat ttgaagggag 480
agtcttcggg agtgataaaa gagaccttgt cctctgtgtg gaggaattat ttagaggcag 540
atgaagttga gcggccgttg agtccgacca gcagcggcta cgctggtgag atggggagca 600
ccagcggcgg tgatgaaatc aatgatgatg acgaagatat taatgaaatt aaagaggtcg 660
aaaacgactt tgtttcgggg tcccatgatc agtgggtttc aggaaaacgc cacccagatg 720
aggatgatgc ttccgtttct tggaggaaaa ggaagaaaca tttctttatc ctgagtcact 780
ctgggaaacc catatattcg agatatggag atgaacacaa gctagctgga ttctctgcaa 840
ctttgcaagc cattatttcc tttgttgaaa atgggggaga ccgggtcaaa ttggttagag 900
ctggtaaaca ccaggtgatt tttcttgtga aaggacccat atacttggtt tgcataagtt 960
gcacggaaga accttatgaa tcactaaaag accagatgga gctactctac ggtcagatga 1020
ttctcatctt gacaaagtct gtaaatcgtt gttttgagaa gaatccaaag tttgatatga 1080
catcattgct ggggggaact gatgctgtct tctcttctct catccattct ttcagttgga 1140
atcctgccac ttttctacat gcctactcat gtcttcccct tgcttatcca acacgtcaag 1200
ccgcaggtgc catcttgcaa gatgtagctg gttcgggagt cctatttgcg cttttgatgt 1260
gtaaacacaa ggttatcagt cttgttggcg cccaaaaagc atctcttcat cctgatgata 1320
tattattact gtcaaatttg ataacgtcct ctgaatcttt taggacatct gaatccttct 1380
caccggtttg cctgccaaga tacaatccca tggcatttct ttatgcttat gtgcattact 1440
ttgatattga tacatacctg atcttgctca ccacaagttc cgatgccttc tatcatttaa 1500
aagattgcag gattcgaatt gaaaacgtac ttttgcagtc aaatgtactg agtgaagttc 1560
aaagatcctt ggtggatggc ggtatgcata tagaggattt gcttgctgac cccacatctc 1620
gtcctggatc cgtttcttct catttaggtc aaccagggcc tggtacaggt tccccggggc 1680
gacttacaga tggatttgtt ggaattggtg gtccagctgg gctatggcat ttcatataca 1740
gaagtattta tcttgaccag tatgtgtcat ctgaattttt atcaccaatc agcagcttga 1800
gacagcaaaa aagattgtat agagcgtacc aaggacttta tgcatccatg catgacaaag 1860
gaattggacc tcacaaaact cagtttagaa gagatgaaaa ttatggtgac atttactaaa 1920
gcaaccaaat tcttcaatta tatgcaagac tggaactctg gaagatagtt tacatcgaga 1980
ttctatgggg tagagccata aattgaagcc atgctactct gctgggtcac acaagatttt 2040
gaactatatg cagcatttga tccactagca gacaaggcac ttgctataaa ggtgtgcaac 2100
cgggtttgtc aatgggtgag agacgttgaa aatgaaatat ttttattggg agtaagccct 2160
ttttcgtggt gattatctca agtattctgt acactggcat gtattctagt ctttccgttc 2220
gactattact taactaccgc ttcctatatg aaattttgcc ttatttttgt tgtacggcct 2280
ctatgttggt ttgcagttcg gatctctatc ctattttttc ttcaccactt gtatctaact 2340
ataaaaacga ataaattgaa gttctatagt gagcttaatt gtagttgttg taattggatg 2400
tttataaaaa ggtaacgttt taacagtgag acataagtgg tatatgggga agtatgattt 2460
tagaggatac cga 2473
<210> 9
<211> 1456
<212> DNA
<213> 海州常山的PROF基因(Clerodendrum trichotomum Thunb)
<400> 9
actaacttga taccaatacg cttgcgatta attgttatct ccgccgagag agaagctctc 60
gcatctttct aagaaagaaa ttttcacttt agatctattt gttttcactc tccatcctca 120
cctccctatc tttccgacgt tgaagtctct tcctcacttc ttgatcctat actaactcca 180
gtttctacat ttgaaatgaa gatggaagat ctttttaggt ctagacgcaa ccagagttgt 240
gtgacagcag cagcggcaag attagggcga ctgcagaagg tggatgtagt tccaccatgg 300
cagtgaccct tgatgtgtgt atttccaaat tttctttagt gtagttttac gttgttttca 360
tgttcatcct cgatctggac tggttctcaa ggtgctagtt ttagggtgct ttatactagc 420
tagtcaagtt gggacagtga cctgaatctc tctctaccaa tgtccttgaa tagggttttg 480
gtccggtcaa gctttcttta ttttgtctcc tttagtacga gtggtttttc ttccatcttt 540
gggattgacc gttcattttg tacatttagt cagttggcta tgtattccac atgtatgtga 600
agttttgggg gaagaagata actaagggga aaaaatttca aagccaattc tgtatttgta 660
aagatgtcca taaaatttga acatctatca gagaaccaac caaaaataca taatttactc 720
acatggaatc ataaacccaa gttttcacaa cgagaaaatc aaaagacaag ccctgtttca 780
attcagatag ggacataatc taacacctct cactcaaaac accacattct ttttgccata 840
accaacaacc cacgctgcaa atcattgaca aaaaaagcag taaaggatgc cgaatttaat 900
aatattaccc aaaaaattat taatttataa ttatataatg aatattgatt attaaatgaa 960
atgatttgga agacttacca atcaatttca gacaccctgt tcaataagat agtcaccaat 1020
cttctcaaca accatgttgc actggcctgc agtcattggt tcatcataga taccgatgat 1080
caaggccaaa ttggtctttt tgacagtggc accaccagat cccttcttcc ctcggatgac 1140
agcatttggt tcaccttgaa tcaccatgta tttagtaccc ccaatgtaca atccagtagg 1200
agctaatgat cccggattat caaaatcatt cataatagct gttacttcct caggcttgaa 1260
ctgagggaag ttggaactct gtgcccaaac gctgccgtca tgaccgacaa ttgcggcgga 1320
ggtgagatgg caaccgtcga cgtcggccat caagtggtca tctatgtagg attgccacga 1380
catcgtttcg ataggcggaa ggaatcaata ccaggaaggt ttggagaatg ggtgaggact 1440
gatgatagcg tgaaaa 1456
<210> 10
<211> 1516
<212> DNA
<213> 海州常山的MDH基因(Clerodendrum trichotomum Thunb)
<400> 10
tttataagaa ttggtaacat ccattggtat atgatattta tctgtcagca aacgggttca 60
ttacatatct agaattgcaa taaccagaac aactcacgta gcataaggta caacatacac 120
taccgcttat tccaagatgt gtgatcttga atctccaaga ttatttcctt acgaaggaga 180
cccccttatc aatgcttcca tccaactctt tctttgcctt ctccagccca accctttcgt 240
attcatttag tggaccaaga gggaagatct cctcgacccc gttacgtcca agccgtaccc 300
ttgaagcaaa gaatggaagt tccgttacct gggaagaaac aaatgcacat tcaacaacac 360
cggaatctcc cctcaagccc ttcaggcaag catcggcaaa tttcactgct gcatatgcca 420
tggaaagagt tgcagagcct gtacccgctt tggcctcaac aacttcagtc ccaccattct 480
ggatgcgaga tgtcagatac tcagtttctt ctttggtgaa agagcatgca ggtgtaatct 540
gcgagaggag aggtaaaatt gtcacaccag catggcctcc tacaactgga acattaactt 600
cccgaggatc tagccccaaa acttcagcca caaacgtatt agctctgact acatcaagca 660
tcgtgactcc aaggagtcgc cttggatcgt aggtcccagc cttcttgaaa acctctgcag 720
caattggaac tgtggagttc actggattac ttattaagtt aacaatggcc ttagggcagc 780
acttagcaac tccttcacaa agtgtcttta ctattcctgc attaatgttg aaaagatcat 840
ccctcgtcat tccaggtttc ctgggaacac cagcaggaat gattacgagg tccatgccag 900
ttagtgcatc cttcaactgg ttttgaccca gaaaaccacg taccacagca ccagtgttca 960
tatggctgat atctgctgta accccaggag tattaactac atcataaaga tgaagcaccg 1020
aaaccagtgg gttcatcttc atcagcattg ctaaaggctg gcctatgccg ccagctgccc 1080
ccaataccgc caccttgaaa cctggtgacc ctccttttgc tcggcaatcg gaaagccata 1140
aactcccctc cacctgcagc tgattagtag gaggatcgag gtgagctgaa attctggaaa 1200
ttcttcgatg aatatcccca ccctgctgca tactataact cgaacgaatt cttctccaat 1260
atcagaaagc agcaaattcc ttttggaata atgagaaatg aaatgatgga agataacttg 1320
aacggcagaa tagtggagat acccgtattt atacggtaga aaattggaat taattgtatt 1380
ttatggcaca gcacataata aaataaataa atacgtcaga ggctcataga aatgcgttaa 1440
tgtatttttg ggtgcatagc cttatcttat acgtacacgc ttcggcaaga aaattacaaa 1500
ttatcaccat aattca 1516
<210> 11
<211> 1351
<212> DNA
<213> 海州常山的EF-1A基因(Clerodendrum trichotomum Thunb)
<400> 11
gaaattcgag accaccaaat attactgcac agtcattgat gcacctgggc atcgtgactt 60
catcaagaac atgattactg gaacctcaca ggctgactgt gccgtcctca tcattgactc 120
cacaacaggt ggttttgaag ctggtatttc caaggatgga cagacccgtg agcatgctct 180
gctggctttc actcttggtg tcaagcaaat gatctgttgt tgtaacaaga tggatgccac 240
tacaccaaaa tactccaaag caaggtatga tgaaattgtg aaggaagtgt cctcttacct 300
caagaaggtt ggatacaacc cggacaagat accatttgtc cccatttctg gttttgaggg 360
agataacatg attgagaggt ctaccaacct agattggtac aagggcccca cgctactcga 420
tgcacttgac cagattaacg agcccaagag gccaacagac aagccactcc gtctcccact 480
tcaggacgtt tacaagattg gtggcattgg tactgttcca gtaggccgtg tggagactgg 540
tgttctaaag cctggtatgg tggtcacctt tggcccgtct ggtctcacca ctgaagttaa 600
atcagttgag atgcaccacg aagccttgca ggaggctctt ccgggtgaca atgtcgggtt 660
caacgtcaag aatgttgctg tcaaggatct aaagcgtggc tttgtggcct ccaactcaaa 720
ggacgaccct gctaaggaag ctgcgaactt cacctctcag gtcatcatca tgaaccaccc 780
tggccagatc ggaagtggct atgcacctgt tctcgactgc cacacctccc acatcgctgt 840
taagtttgct gagttagtca ccaagattga taggcgttcg ggtaaggaga ttgagaagga 900
gccaaaattc ttgaaaaatg gtgatgctgg attcgtgaag atggttccga ctaagcctat 960
ggttgtcgag actttctctg agtacccacc tctgggtcgt tttgctgtac gagacatgcg 1020
acagactgtt gctgttggcg ttattaagag cgtggagaag aaggacccga gtggtgctaa 1080
ggtgaccaaa gctgctgcta agaagggtgc taagtgaacg ctggtgactc cgttttatca 1140
ataaattttg tttttataat gcagtttatg attaatgatt attgctttta ctgctgtatc 1200
tctagcctct tgtaagagga tattggagag tggagcaact tggttttgat tcttgttgct 1260
tcatgtgcag tgatgaaagc agctctgttt ggatattgtc aaatttttat tttccttgtg 1320
gattttgttt tcttgaaatt gttacttagt g 1351
<210> 12
<211> 1214
<212> DNA
<213> 海州常山的UBC-E2基因(Clerodendrum trichotomum Thunb)
<400> 12
aaattgtttt gcaaaaacct cgattttcat agtactcgtt cgtcggtgtg cgatctgagg 60
agatggcgga agataagtat aacaggaaga accctgctgt gaagaggatt ttgcaggagg 120
tgaaagagat gcaatccaat ccttccgatg atttcatgag ccttcctctc gaggagaaca 180
tatttgaatg gcagttcgca ataagagggc caagagactc ggagttcgag ggaggaatat 240
atcatggaag aattcagttg cctgctgaat acccctttaa gccaccttca tttatgttat 300
tgacgccaaa tggtagattt gaaacccaaa ccaagatatg cttgagcatt tcaaatcatc 360
accctgaaca ctggcaacca tcctggagtg ttcgaactgc gctggttgca ttgattgcat 420
ttatgcctac tagcccaaat ggtgcgctag gatctttgga ttacacaaaa gaagagaggc 480
gagctcttgc agtaaaatct cgcgaagcag ctccaaaatt tggatcacct gaaaggcaaa 540
ggctgattga tgagattcat gagtatatgc tcagcaaagc gctgcctgtg cctcaaggca 600
acctcacctc tgaagcacac actgggagtg aaaaaatcca gcagaccacc caagacaatg 660
ttgaggaagt tacagaagca cctccccaca cagctgaaga cgacgggatc atacaagaac 720
caagtgaagc agtagcacct tctactaata caaatgttgc tcagacatcg cagcctgttc 780
ctcccgggcc gacgacgagc cagcagcagc tgttgcataa gcccgaacca agggttccta 840
aaccagcaga tgatcgtctc ttcacatggg ctgctttcgg gctcaccata gctatagtag 900
ttcttctgtt gaagaagttc ttgaaggcta atgggcacgg tgctgtcttt atgaatgagt 960
cgtaagtggg acctggtatg ttgtttatgc tgatgcagaa ggtgaatgag gggaaaacct 1020
gctttctgta atggagtgcc ttaaatttta aggggaacat atctatatat atattatacg 1080
tatacttatc tttctactta tggagtatat tgtatagttt aaataattgt taggggtaat 1140
gttctgaagc tttttaagaa gctgaactat tttaatagaa gcagttctgt aattattgct 1200
tgcatttgtg agag 1214
<210> 13
<211> 2673
<212> DNA
<213> 海州常山的AP-2基因(Clerodendrum trichotomum Thunb)
<400> 13
ttaatcaccg atgtcttcag aaccaatacg caactaattg acaaacagat gctgtaattg 60
ttacaaaggg tgcagttttc attatgtcac ttgcattaca gaaaccaatg aagaagggtg 120
tatttggcaa gcgctaaata aagaggaaaa aaaatctcac caaggcacaa tctgattgtg 180
cgaaagaatt atcaaaactt caaatccaaa ttctaacact atgccgatta catacaaaat 240
tacaaattca ccctccgact cccccagctc ttgcaccata ttagcgggga tccctccagt 300
acatacgagc aggtgctata tgaacattgg ttaaacaatt ttgaacagcc ttgggtttgt 360
tttatcactc aaagcaaacc agcaagaata gctccaggat ctgatgtaga tgtaggtgga 420
ggcgttagtt gtggttgtgg ttgaggttga ggttgaggtt gaggttgagg tggcgtaggt 480
gcaggtgctc gagaagggat gggaatggtg attaattgtt ctttaatgaa ttctttcaat 540
tcgtatgtta gttttggatc tccagatgcg atggtcatcc gtaattgcgt tctatctgct 600
gggtctgtct cgattcttat caaacataac atggctcgtg tactttcaga gtagaaagtt 660
gtgctcgcaa ccaaattatt agcatttgga tcaagcccag ggcaaaccat caggtgcaaa 720
ctgctaaata agtttgccat ctctgctagc agcattggtc ttacacctct aaccacttct 780
tgcagtttca atggaggtcc tgatagtgat ctccattgtg ggaaaaattc ttcagcagaa 840
actgtaattg actgaagaaa tttgttcaaa acagcaggaa ggcgaagttt tacattgacc 900
aaatgggttc caaacttgta cgagaagtcc agtactgcta aatctctgct cggttggagg 960
atgattactt caagtgggca ttgaacctgt gcccttggag gaatagtctc aggtactaaa 1020
gatagttcca tctttagatg tgatggaggc aatattaaag cttgaactga aacgagtgga 1080
gcagtatttt tatttcccaa gaagagaaca actcgcccat gatgggttcg ccattcagct 1140
tttatgccaa tctggatata ggcatcttca tatagcacac cactatcctt caggcacaaa 1200
acacgaaatc tttccactat atcaccaatc ggctggaccg tattaacttg ctcttcaaca 1260
ggtgctatgg ctaatgcatc tgtttccaaa ccggaagcaa cccttggatc agtttgggga 1320
gcaatgcctg gtgggccctc aatagccagt ggccctaaaa gatctccaag gagatcagga 1380
gaaggtgcac tgtgtggttg aggatccact acactcaggg caccattagc acgagtcaac 1440
ccttgctcag cagtgctctg atccgcatta ctcatactag gcaccttgac aagaccaagc 1500
tgggtcccag atggtgttcc atttgcaggg cgttgatgat cggtaagtac caaagcatta 1560
gaattctgct gctgtgcccg taattttatt gcactttgct cagcagtatc agcttcagca 1620
tcttccgctt tttttatcaa tgatgactgt ctctcaggga atttcggcat ttcagcaagt 1680
atatccacta gagcagcacc tttcatgctc agagcaagat actcaacagc tcgttgctgt 1740
atttcagcat caatgcaact ttcatacttt ttgaatatca cccatatttg cttctgtagc 1800
tcctggctgg gtggttgagt gtgcatcaaa atttttgcat atgtggaaag aaggatagga 1860
attgttgatg tcgagacagt gggaagcttc tcatgtatca agttgaaaat ttcctttgga 1920
ctgcacccag gtcgcctagc tagaagatga ctgtattctc caaggatata tgcactcacc 1980
ttcaccatgg tctcgtgaat cgcaggctta tcaagatact ctttggcttt cagagctgca 2040
taaggctgta aatcctcgtt gtttgtaacg aactgcacga ctcgaaacca aatgtcatca 2100
ccgacaaaat cccctgcctt gtcaattagc tgaagaatca catccacata ccatgacagg 2160
tcgggggcaa acttctctgc gagaatagca gctttaagtg acaattcttc ccgcatagca 2220
aagtcagctg agctcagata ctgtaataat tcttcaacta tgtcctttgc atttgaaaca 2280
tcacacattc catatagcaa atcaagggca cgccttctga tactgatatc aggatccttc 2340
agggaagtaa tgatctgagc ttgatgtctt ttgataattt catgcacatc ggcgaccatc 2400
aacattcgag tcatgttctc caagccaagg tatcgtatat taggttcacg aacagcaata 2460
aattttccca gcaaagcaac acactgagac atcatttctt tctctgcatc aagatgcatg 2520
acaagagcaa gagcttcaaa aagaacagca tgtgatgcat tattcttgtt cacatttttc 2580
accacatcag ttcccatgag tatccgttgc agaacctcaa acaatgatct tcttgtattt 2640
ggatcctcaa ctgttggaaa atactggaga gcc 2673
<210> 14
<211> 2071
<212> DNA
<213> 海州常山的HSP70基因(Clerodendrum trichotomum Thunb)
<400> 14
tgttagtttt atttcattat tgttctttga cttgagcaaa attctctggt cgatttagat 60
ctgcgattag ttttgcgagc aatggccgga aaaggagaag gtccagcgat cgggattgat 120
ctcggaacga cgtactcttg tgttggcgtt tggcaacacg accgtgttga aatcatcgcg 180
aatgaccagg gtaacagaac gacgccgtct tatgttggct tcaccgattc agagaggctt 240
attggtgatg ccgctaaaaa tcaggtcgcc atgaacccga cgaacactgt ttttgatgcc 300
aagaggttga ttggaaggcg atttagcgat gcttctgttc aaagcgacat caaactatgg 360
cccttcaagg tcattgccgg acctggtgag aaacccatga ttgttgtcaa ctataagggt 420
gaagacaagc agtttgcagc tgaggagatt tcatctatgg tcctgatgaa aatgaaggaa 480
atagcagagg cctaccttgg ttccacaata aagaatgctg tcgtcaccgt tcctgcatac 540
ttcaacgact cccagcgtca agccaccaaa gacgctggcg taatttcagg tctcaatgtt 600
atgcgtatca tcaacgaacc tacagctgcc gccattgctt atggtcttga caagaaggcc 660
tcgagtgtag gtgagaagaa tgtcttgatt tttgatcttg gtgggggcac ttttgacgtc 720
tctctgttga cgatcgagga gggtattttc gaagtgaagg ccactgctgg cgacacccat 780
cttggaggtg aggactttga caacagaatg gtgaaccact ttgttcagga attcaagagg 840
aagagcaaga aggatatcac cggtaacgct agagctctac gaaggctgag aactgcttgt 900
gagagagcaa agagaaccct ttcatccaca gcccaaacaa caatcgagat agactctcta 960
tacgaaggaa tcgatttcta ctccaccatc acccgtgcaa ggttcgagga gcttaacatg 1020
gatctgttcc gcaagtgtat ggaaccagtt gagaagtgcc tgcgcgatgc caagatggac 1080
aagagcacgg tgcatgacgt tgtgctcgtc ggtgggtcca cccgtatccc caaggtgcaa 1140
cagctgcttc aggacttctt caacgggaag gagctttgca agagcatcaa cccggatgaa 1200
gccgtggcct acggcgccgc cgtccaagcc gccattttga gtggtgaggg taacgagaag 1260
gttcaagacc tattgcttct cgatgtcact cccctctccc ttggtttgga aactgccgga 1320
ggtgtaatga ctgttttgat tcccaggaac accaccattc ctactaagaa agagcaggtg 1380
ttctctacct actcggacaa ccaacccggc gtcttgatcc aggtctacga gggtgagcgt 1440
acgaggacga gggacaacaa cctgctcggg aagtttgagc tctccggcat tccccccgcc 1500
cctcgtgggg tcccgcagat cactgtgtgc ttcgacatcg atgctaacgg cattctaaac 1560
gtgtcggccg aggacaagac gacgggccaa aagagcaaga tcacaattac aaacgacaag 1620
gggaggctgt caaaagacga gatagagaag atggtacagg aggcggagaa gtacaagtcg 1680
gaggacgagg agcataagaa gaaggtggaa gcgaagaatg cgctggagaa ctatgcgtac 1740
aacatgagga acacggttaa ggatgaaaag atcgggtcga agctgcccgg tgctgataag 1800
aagaagattg aggatgcggt tgagtcgacg attcaatggc tcgatgggaa ccagctcgcg 1860
gagtcggatg agtttgagga taagatgaag gagttggagg gcatttgtaa cccgattatt 1920
gccaagatgt atcagggtgc ggggggtgcg gcggctatgg atgacgatga cggtctggcg 1980
gcggcgcctg gtggtgctgg tcctaagatt gaagaggttg actaagttgt ggcccctttt 2040
ttggtggttt tttttttttt tttttttttt g 2071
<210> 15
<211> 1802
<212> DNA
<213> 海州常山的TUA基因(Clerodendrum trichotomum Thunb)
<400> 15
atctgagccc tagagagaga aagtgaaatt acaccttcat tttctaccaa ctgctcttat 60
aaaatcccct ttcaccacga attaagaact acctattttc atccagagac gtcttcgtat 120
acgccctggt atttctctcc tttgatctcc gatcgaaatc catttcagat tttaatcttt 180
caaacaaaaa gcgaaaatga gagagtgcat ttcgatccac attggacaag ccggaatcca 240
ggtcggcaat gcctgttggg agctgtactg ccttgaacat ggcatccagc ctgatggaca 300
aatgcctggt gatcacactg tcggaggagg tgatgatgcc ttcaacacat tcttcagcga 360
aactggcgcc gggaagcatg ttccccgtgc agtgttcgtc gatctggagc ctactgtgat 420
tgatgaggtg cgcactggta cataccgtca gctattccac cctgaacaac taatcagcgg 480
caaagaagac gctgccaaca actttgccag aggtcattac actattggca aagaaatcgt 540
ggatctgtgc cttgatagga tccgcaaact cgctgacaac tgtactgggt tgcaaggctt 600
cctcgttttc catgctgtgg gtggtggtac cggatctggc ctcggatctc tgttgctcga 660
gaggctctcc gtcgattatg gtaaaaagtc gaagctgggt ttcactatct acccttcccc 720
tcaagtctca actgccgttg ttgagcctta taactctgtg ctttcaactc attcccttct 780
tgaacacact gatgttgctg ttcttcttga taatgaggcc atctatgata tttgccgcaa 840
atctctggac atcgagaggc ctacatacac caatttgaac aggctcattt ctcaggtgat 900
ttcctcctta actgcatccc tgagatttga cggagcattg aacgtcgatg taaacgaatt 960
ccaaaccaac ctcgtcccat acccaagaat ccacttcatg ctttcatcat acgcccccgt 1020
aatctcagct gagaaagcct accacgagca gctctccgtc gctgagatca ccaacaccgc 1080
ctttgagcca tcttccatga tggtgaagtg cgatccgcgc cacggcaagt acatggcgtg 1140
ctgcctcatg tacagaggcg atgttgtccc caaagatgtg aacgcggccg ttgccacaat 1200
caagacaaag aggacgatcc agtttgtcga ctggtgccca actggcttca agtgtggcat 1260
caactaccag ccgccgactg tggtccctgg tggtgatctt gctaaggtgc agagggcagt 1320
ttgcatgatt tcaaactcga ctagtgttgc tgaggtgttc tcgaggatcg atcataagtt 1380
tgatttgatg tatgctaaga gggctttcgt gcattggtat gtgggtgagg gtatggagga 1440
aggggagttc tcggaggcga gggaggattt ggctgctttg gagaaggatt atgaggaagt 1500
tggggctgaa tctgctgaag gtgaagatga tgagaatgaa gagtactaga gtagggacca 1560
attggctcgt tggtttagtg ctgaactttt ttccttatgg ttgaacttct tgtctggtgt 1620
tgaactgctt atggattctc tgtgtgtagt tcgtattgat tatggtatgt gttgaaaagg 1680
ggctttatgt tcatttcgta ttgattatgc ttgtgtaatt cgtatgtgtt gattattagt 1740
tattccctag gtcatactcg tattttatat gtttaaaatg ccaagataat tttttttttt 1800
tt 1802
<210> 16
<211> 1670
<212> DNA
<213> 海州常山的UBQ基因(Clerodendrum trichotomum Thunb)
<400> 16
gaattgttct cataaatttt cagttttttc gtctctcaat cgtcaagatg caaatcttcg 60
tgaaaactct caccggcaag accatcaccc ttgaggttga aagctccgat accattgaca 120
acgtcaaggc taagatccag gacaaggaag gtatcccccc agaccagcaa aggctgatct 180
tcgcaggaaa acagcttgag gatggccgta ccctcgccga ctacaatatt cagaaggaat 240
caactctcca cctggtcctc cgtctccgtg gaggtatgca gatattcgtg aagaccctaa 300
cgggcaagac catcaccctc gaggttgaaa gctccgacac gatcgataac gtgaaggcca 360
agatccagga caaggagggt attcccccag accagcagag gctgatcttc gccggcaagc 420
agctcgagga cggacgtacc ctcgccgact acaatatcca gaaggagtcc accctccatt 480
tggtcctccg tctccgtggt ggtatgcaga tctttgtcaa gaccctcacc ggcaagacca 540
tcaccctcga ggttgaaagc tccgacacga tcgataacgt gaaggccaag atccaggaca 600
aggagggtat tcccccagac cagcagaggc tgatcttcgc tgggaagcaa ctcgaggatg 660
gacgcacgct tgctgattac aacatacaaa aggaaagcac acttcatttg gttctgaggt 720
tgagaggtgg aatgcaaatc tttgtcaaga ctctcacggg gaagaccatc accttggagg 780
ttgagagttc cgacaccatt gataatgtga aagccaagat acaagacaag gaaggtatcc 840
caccagacca gcagaggctc atatttgcag ggaagcagct ggaggatgga cgcaccttgg 900
ctgattacaa cattcagaag gagtctactc ttcatttggt gttgaggttg aggggaggga 960
tgcagatatt tgtcaagaca ttgactggga agaccatcac tttggaggtg gagagttcag 1020
acactattga taatgtgaag gccaagatcc aggataagga gggcatccca cccgaccagc 1080
agaggttgat cttcgccggg aaacagctgg aggatgggag gactctcgct gattacaata 1140
tccagaagga atccaccctt caccttgtcc tcaggctccg tggtggtttc tagagcttct 1200
ggttttcttg ttgtttgaat gttctggttt tatgttctgt ttgttgattt tacatgattc 1260
ctgttttccc gtgtgtgtgt gcgtgtgttg aaacttaagt atggtactgt tatgttgtcg 1320
tcttgctgac attctcagtt gtcattggaa taaaattggg attttatccc ccctataata 1380
aatttccggt gagtgcataa ttggctgtaa tagttttttc aatattgttg ctgttgagaa 1440
ggtgctacgt aatgttctac cttcttcatc ggttgctttt ctggagattc ttgtatgcct 1500
cggtaatgtt tggtgattat atttatcttc tttaattact gaaagaagtt ggatccgatg 1560
cactatccca ggagaggtcg aagcgatgta ttcctaggtg tttttctgtt agtggagttt 1620
acttcgttgc atgttgtccc gtcaaccatt ttcgtaaggg attaatgttt 1670
<210> 17
<211> 588
<212> DNA
<213> 海州常山的H3基因(Clerodendrum trichotomum Thunb)
<400> 17
actgatcagt tttactacaa aacagacaaa ggttaaacat tcaaagccac aagtgacacc 60
tcgaaacaac cacatattta ccatcggaaa aaactatccc ttagacccaa atcatccaga 120
ttacaagtta agccctttca cccctaattc tcctcgcaag ctggatatct ttgggcataa 180
ttgtaaccct cttagcatga atggcacaga ggttagtatc ctcaaacaat ccaacaagat 240
aggcctccgc tgcctcttga agagccgcca cagcagagct ctggaacctc agatccgtct 300
tgaaatcctg agcaatctca cggacaagcc tctggaaagg aagttttcgg atcaaaagct 360
cagtggattt ctggtacttg cgaatttcac ggagagcgac ggttcccggg cgaaaacggt 420
gaggcttctt cactccgccg gtggcggggg ctgatttcct agcggctttg gtggccaact 480
gcttccgcgg tgccttgcct ccggtggatt tgcgggcggt ttgcttagtg cgagccattt 540
ctagggtatc gatagaaagt atggaaaaaa agtgctaatt tggaattg 588
<210> 18
<211> 514
<212> DNA
<213> 海州常山的ClNHX1基因(Clerodendrum trichotomum Thunb)
<400> 18
gccaataatt tctcagcaat ggagggctta ttttaccgtc ttctccttct tacagttcca 60
ttaatggcca tgcaagtgct tccttcttct tcatcatcca atacaaagat gtttcagttt 120
aatgttgaat ggaagagtat aacaagactt tgcaatagaa agacagtatt gacagtaaat 180
gggaagtacc cagggccaac cattgctgtg aatgaaggtg acagtgttca agtaacggtc 240
actaattggg ttgcaagaaa caccaccatc cattggcatg gagtaaggca gatgagaagg 300
ggttgggcag atggggcagc atacataaca caatgcccga ttggaggagg caaaagctat 360
agatacaact ttagtgtaga ggatcagagg ggtacactgt ggtggcatgc tcatatcggc 420
tggcaacgtg cttcggttta tggcgctttt atcatttacc ctcgcacgcc ttttccattc 480
tctctccaac tccacccact tcagattcct ctca 514
<210> 19
<211> 21
<212> DNA
<213> Actin引物序列(Artificial)
<400> 19
cgattcttgc ttccctcagt a 21
<210> 20
<211> 18
<212> DNA
<213> Actin下游引物序列(Artificial)
<400> 20
accattttcg ccgcctta 18
<210> 21
<211> 19
<212> DNA
<213> PP2A上游引物序列(Artificial)
<400> 21
ggacaagcac caagacgcc 19
<210> 22
<211> 22
<212> DNA
<213> PP2A下游引物序列(Artificial)
<400> 22
tcagcacttc taaaaccgca tc 22
<210> 23
<211> 21
<212> DNA
<213> RPL上游引物序列(Artificial)
<400> 23
agtcaatggt ggcgatgtag c 21
<210> 24
<211> 22
<212> DNA
<213> RPL下游引物序列(Artificial)
<400> 24
cccttggtca ctccgataat gt 22
<210> 25
<211> 23
<212> DNA
<213> PK上游引物序列(Artificial)
<400> 25
ctgaaggtgg tgtactcctg ctc 23
<210> 26
<211> 21
<212> DNA
<213> PK下游引物序列(Artificial)
<400> 26
cgaaactgcc ccaacaagat a 21
<210> 27
<211> 21
<212> DNA
<213> 18S上游引物序列(Artificial)
<400> 27
acaatcaccg aacaaagcag c 21
<210> 28
<211> 22
<212> DNA
<213> 18S下游引物序列(Artificial)
<400> 28
cgagaagtaa tcttggaacg cc 22
<210> 29
<211> 20
<212> DNA
<213> RAN上游引物序列(Artificial)
<400> 29
ggagacgaca gtttgggtgc 20
<210> 30
<211> 19
<212> DNA
<213> RAN下游引物序列(Artificial)
<400> 30
tgccatactt gcgggtgaa 19
<210> 31
<211> 19
<212> DNA
<213> APT上游引物序列(Artificial)
<400> 31
ccacgcattc aagccattc 19
<210> 32
<211> 20
<212> DNA
<213> APT下游引物序列(Artificial)
<400> 32
gagcctctat cccagcaacg 20
<210> 33
<211> 19
<212> DNA
<213> SAND上游引物序列(Artificial)
<400> 33
gaccagtcgc ttgatgccc 19
<210> 34
<211> 22
<212> DNA
<213> SAND下游引物序列(Artificial)
<400> 34
ggatgctcca ggtactgctc tt 22
<210> 35
<211> 24
<212> DNA
<213> PROF上游引物序列(Artificial)
<400> 35
gaatgtggtg ttttgagtga gagg 24
<210> 36
<211> 18
<212> DNA
<213> PROF下游引物序列(Artificial)
<400> 36
atgtgaagtt ttggggga 18
<210> 37
<211> 20
<212> DNA
<213> MDH上游引物序列(Artificial)
<400> 37
acctggaatg acgagggatg 20
<210> 38
<211> 24
<212> DNA
<213> MDH下游引物序列(Artificial)
<400> 38
gccacaaacg tattagctct gact 24
<210> 39
<211> 20
<212> DNA
<213> EF-1A上游引物序列(Artificial)
<400> 39
aggatggaca gacccgtgag 20
<210> 40
<211> 22
<212> DNA
<213> EF-1A下游引物序列(Artificial)
<400> 40
aaaaccagaa atggggacaa at 22
<210> 41
<211> 23
<212> DNA
<213> UBC-E2上游引物序列(Artificial)
<400> 41
gcaaaggctg attgatgaga ttc 23
<210> 42
<211> 21
<212> DNA
<213> UBC-E2下游引物序列(Artificial)
<400> 42
cctcaacatt gtcttgggtg g 21
<210> 43
<211> 20
<212> DNA
<213> AP-2上游引物序列(Artificial)
<400> 43
ccacaactaa cgcctccacc 20
<210> 44
<211> 20
<212> DNA
<213> AP-2下游引物序列(Artificial)
<400> 44
aaattcaccc tccgactccc 20
<210> 45
<211> 20
<212> DNA
<213> HSP70上游引物序列(Artificial)
<400> 45
gccattttga gtggtgaggg 20
<210> 46
<211> 23
<212> DNA
<213> HSP70下游引物序列(Artificial)
<400> 46
ggttggttgt ccgagtaggt aga 23
<210> 47
<211> 20
<212> DNA
<213> TUA上游引物序列(Artificial)
<400> 47
gtccccaaag atgtgaacgc 20
<210> 48
<211> 20
<212> DNA
<213> TUA下游引物序列(Artificial)
<400> 48
aagcagccaa atcctccctc 20
<210> 49
<211> 21
<212> DNA
<213> UBQ上游引物序列(Artificial)
<400> 49
tgctgttgag aaggtgctac g 21
<210> 50
<211> 19
<212> DNA
<213> UBQ下游引物序列(Artificial)
<400> 50
cgaaaatggt tgacgggac 19
<210> 51
<211> 21
<212> DNA
<213> H3上游引物序列(Artificial)
<400> 51
caagttaagc cctttcaccc c 21
<210> 52
<211> 21
<212> DNA
<213> H3下游引物序列(Artificial)
<400> 52
tgctcaggat ttcaagacgg a 21

Claims (8)

1.海州常山干旱胁迫下荧光定量内参基因，其特征在于，为Actin、AP-2和RAN基因，其中，Actin基因的基因序列如SEQ ID NO.1所示，AP-2基因的基因序列如SEQ ID NO.13所示，RAN基因的基因序列如SEQ ID NO.6所示。
2.权利要求1所述的海州常山干旱胁迫下荧光定量内参基因的专用引物，其特征在于，Actin基因的引物序列如下：
Actin正向引物5’-CGATTCTTGCTTCCCTCAGTA-3’
Actin反向引物5’-ACCATTTTCGCCGCCTTA-3’。
3.权利要求1所述的海州常山干旱胁迫下荧光定量内参基因的专用引物，其特征在于，AP-2基因的引物序列如下：
AP-2正向引物5’-CCACAACTAACGCCTCCACC-3’
AP-2反向引物5’-AAATTCACCCTCCGACTCCC-3’。
4.权利要求1所述的海州常山干旱胁迫下荧光定量内参基因的专用引物，其特征在于，RAN基因的引物序列如下：
RAN正向引物5’-GGAGACGACAGTTTGGGTGC-3’
RAN反向引物5’-TGCCATACTTGCGGGTGAA-3’。
5.权利要求1所述的海州常山干旱胁迫下荧光定量内参基因在海州常山荧光定量中的应用。
6.权利要求2所述的Actin基因的引物序列在海州常山荧光定量中的应用。
7.权利要求3所述的AP-2基因的引物序列在海州常山荧光定量中的应用。
8.权利要求4所述的RAN基因的引物序列在海州常山荧光定量中的应用。
CN201811441119.7A 2018-11-28 2018-11-28 海州常山干旱胁迫下荧光定量内参基因及其专用引物和应用 Expired - Fee Related CN109207634B (zh)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110016516A (zh) * 2019-04-12 2019-07-16 中国热带农业科学院热带生物技术研究所 狗尾草响应不同干旱及盐胁迫表达的荧光定量内参基因Actin及其应用
CN110079627A (zh) * 2019-04-25 2019-08-02 四川天艺优境环境科技有限公司 用于小报春不同花期基因表达分析的内参基因及其引物
CN111826424A (zh) * 2020-07-15 2020-10-27 扬州大学 芍药干旱胁迫下实时荧光定量pcr内参基因及其专用引物和应用

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110016516A (zh) * 2019-04-12 2019-07-16 中国热带农业科学院热带生物技术研究所 狗尾草响应不同干旱及盐胁迫表达的荧光定量内参基因Actin及其应用
CN110016516B (zh) * 2019-04-12 2022-11-01 中国热带农业科学院热带生物技术研究所 狗尾草响应不同干旱及盐胁迫表达的荧光定量内参基因Actin及其应用
CN110079627A (zh) * 2019-04-25 2019-08-02 四川天艺优境环境科技有限公司 用于小报春不同花期基因表达分析的内参基因及其引物
CN111826424A (zh) * 2020-07-15 2020-10-27 扬州大学 芍药干旱胁迫下实时荧光定量pcr内参基因及其专用引物和应用
CN111826424B (zh) * 2020-07-15 2021-04-06 扬州大学 芍药干旱胁迫下实时荧光定量pcr内参基因及其专用引物和应用

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