本产品包含两份质粒:J6V-HM(JA生物传感器)和mJ6V-HM(JA生物传感器对照)。
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更多资讯可参阅推文:https://mp.weixin.qq.com/s/RY4aHGf1qqMKu840ZLw4Xw
或者阅读原文献:https://onlinelibrary.wiley.com/doi/10.1111/pbi.13718
J6V-HM生物传感器:含有该构建的植株可以同时表达JA响应元件和核内参元件,通过比较两个元件所带荧光蛋白的比值来推算不同条件、不同细胞组织和发育过程中JA含量、或响应变化的相比值,在时空上快速获得JA的含量或反应的示踪图谱。注意:Jas功能域是抑制蛋白功能DII功能域,高VENUS信号代谢低JA含量。
构建简要信息:
茉莉素感应元件:Jas-VENUS-NIL-6HA,其中Jas功能域为OsJAZ6基因中的Jas功能域
内参元件:H2B-mCherry
启动子:Ubiquitin promoter
蛋白独立翻译短肽F2A:来源于病毒的短肽(〜18-25个氨基酸),它们通常被称为“自我剪切”肽,能使一条转录产物产生多种蛋白。2A肽并不是完全的“自我剪切”,而是通过使核糖体跳过2A元件C-末端的甘氨酸和脯氨酸肽键的合成而发挥作用,最终导致2A序列末端和下游产物分离。
载体:pTCK303-6HA
mJas6:如图所示,在Jas6功能域中进行了RK至AA的突变。该构建是负对照。
质粒抗性:Kanr;
转化植株搞性:Hgyr
Ubi-1:Jas6-VENUS-6HA:F2A:H2B-mCherry 序列信息:
ctgcagtgcagcgtgacccggtcgtgcccctctctagagataatgagcattgcatgtcta
agttataaaaaattaccacatattttttttgtcacacttgtttgaagtgcagtttatcta
tctttatacatatatttaaactttactctacgaataatataatctatagtactacaataa
tatcagtgttttagagaatcatataaatgaacagttagacatggtctaaaggacaattga
gtattttgacaacaggactctacagttttatctttttagtgtgcatgtgttctccttttt
ttttgcaaatagcttcacctatataatacttcatccattttattagtacatccatttagg
gtttagggttaatggtttttatagactaatttttttagtacatctattttattctatttt
agcctctaaattaagaaaactaaaactctattttagtttttttatttaataatttagata
taaaatagaataaaataaagtgactaaaaattaaacaaataccctttaagaaattaaaaa
aactaaggaaacatttttcttgtttcgagtagataatgccagcctgttaaacgccgtcga
cgagtctaacggacaccaaccagcgaaccagcagcgtcgcgtcgggccaagcgaagcaga
cggcacggcatctctgtcgctgcctctggacccctctcgagagttccgctccaccgttgg
acttgctccgctgtcggcatccagaaattgcgtggcggagcggcagacgtgagccggcac
ggcaggcggcctcctcctcctctcacggcaccggcagctacgggggattcctttcccacc
gctccttcgctttcccttcctcgcccgccgtaataaatagacaccccctccacaccctct
ttccccaacctcgtgttgttcggagcgcacacacacacaaccagatctcccccaaatcca
cccgtcggcacctccgcttcaaggtacgccgctcgtcctccccccccccccctctctacc
ttctctagatcggcgttccggtccatggttagggcccggtagttctacttctgttcatgt
ttgtgttagatccgtgtttgtgttagatccgtgctgctagcgttcgtacacggatgcgac
ctgtacgtcagacacgttctgattgctaacttgccagtgtttctctttggggaatcctgg
gatggctctagccgttccgcagacgggatcgatttcatgattttttttgtttcgttgcat
agggtttggtttgcccttttcctttatttcaatatatgccgtgcacttgtttgtcgggtc
atcttttcatgcttttttttgtcttggttgtgatgatgtggtctggttgggcggtcgttc
tagatcggagtagaattctgtttcaaactacctggtggatttattaattttggatctgta
tgtgtgtgccatacatattcatagttacgaattgaagatgatggatggaaatatcgatct
aggataggtatacatgttgatgcgggttttactgatgcatatacagagatgctttttgtt
cgcttggttgtgatgatgtggtgtggttgggcggtcgttcattcgttctagatcggagta
gaatactgtttcaaactacctggtgtatttattaattttggaactgtatgtgtgtgtcat
acatcttcatagttacgagtttaagatggatggaaatatcgatctaggataggtatacat
gttgatgtgggttttactgatgcatatacatgatggcatatgcagcatctattcatatgc
tctaaccttgagtacctatctattataataaacaagtatgttttataattattttgatct
tgatatacttggatgatggcatatgcagcagctatatgtggatttttttagccctgcctt
catacgctatttatttgcttggtactgtttcttttgtcgatgctcaccctgttgtttggt
gttacttctgcaggtcgactctagaggatcatggatctgcctcaggctaggaaggcgtcg
cttcaccggttcctcgagaaaagaaaggatcgccttcaggctaaagcaccctactgcggc
agctgcgacccagctttcttgtacaaagtggccgcagctgccgcaatggtgagcaagggc
gaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggc
cacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctg
aagctgatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctg
ggctacggcctgcagtgcttcgcccgctaccccgaccacatgaagcagcacgacttcttc
aagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggc
aactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgag
ctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaac
tacaacagccacaacgtctatatcaccgccgacaagcagaagaacggcatcaaggccaac
ttcaagatccgccacaacatcgaggacggcggcgtgcagctcgccgaccactaccagcag
aacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccag
tccgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtg
accgccgccgggatcactctcggcatggacgagctgtacattgctgcagcggccgaattc
aagcgtgaagagcaagcaaggaaagctaaggtgaacaatgagaaaaagacggaaatagtg
aaaccagagagttgtagcaatgaaggagatgtcaaggatctgaaaagaaaggactctgag
gatggaaacgagggtgaggaagaagaagcttcttcgaaaccgaaaaagccaaaagttgct
ctttctcatcttcaggacattgacgacacagaagctgatcaagaagaagagggtaccatg
ggaagatctactagttctagttacccatacgatgttcctgactatgcaggctatccctat
gacgtcccggactatgcaggttcctatccatatgacgttccagattacgcttcaagatac
ccatacgatgttcctgactatgcgggctatccctatgacgtcccggactatgcaggttcc
tatccatatgacgttccagattacgctactagagtgaagcagaccctcaacttcgacctc
ctcaagctcgccggcgacgtggagtccaatcctggcccgggatccatggcgcccaaggcg
gagaagaagccggccgcgaagaagcccgcggaggaggagcccgcggcggagaaggccgag
aaggccccggcggggaagaagcccaaggcggagaagcgtctccccgccggcaagggcgag
aagggcagcggcgaggggaagaaggcggggcggaagaaggggaagaagagcgtcgagacc
tacaagatctacatcttcaaggtgctcaagcaggtccaccccgacatcggcatctcctcc
aaggccatgtccatcatgaactccttcatcaacgacatcttcgagaagctcgccgccgag
gccgccaagctcgcccgctacaacaagaagcccaccatcacctcccgggagatccagacc
gccgtccgcctcgtcctccccggcgagcttgccaagcacgccgtctccgagggcaccaag
gccgtcaccaagttcacttcctctgatccaccggtcgccaccatggtgagcaagggcgag
gaggataacatggccatcatcaaggagttcatgcgcttcaaggtgcacatggagggctcc
gtgaacggccacgagttcgagatcgagggcgagggcgagggccgcccctacgagggcacc
cagaccgccaagctgaaggtgaccaagggtggccccctgcccttcgcctgggacatcctg
tcccctcagttcatgtacggctccaaggcctacgtgaagcaccccgccgacatccccgac
tacttgaagctgtccttccccgagggcttcaagtgggagcgcgtgatgaacttcgaggac
ggcggcgtggtgaccgtgacccaggactcctccctgcaggacggcgagttcatctacaag
gtgaagctgcgcggcaccaacttcccctccgacggccccgtaatgcagaagaagaccatg
ggctgggaggcctcctccgagcggatgtaccccgaggacggcgccctgaagggcgagatc
aagcagaggctgaagctgaaggacggcggccactacgacgctgaggtcaagaccacctac
aaggccaagaagcccgtgcagctgcccggcgcctacaacgtcaacatcaagttggacatc
acctcccacaacgaggactacaccatcgtggaacagtacgaacgcgccgagggccgccac
tccaccggcggcatggacgagctgtacaagtaa