Duckweed expression vector of neurotransmitter gamma-aminobutyric acid fluorescent probe iGABASnFR and construction method of duckweed expression vector
A fluorescent probe and aminobutyric acid technology, applied in the field of bioengineering, can solve the problem of ecological restoration of heavy metal-polluted water environment and the application of plant resistance to heavy metals. The physiological mechanism and molecular mechanism of duckweed enriching heavy metals have not been clarified, etc question
Pending Publication Date: 2021-11-19
TIANJIN NORMAL UNIVERSITY
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However, the physiological and molecular mechanisms of duckweed enriching heavy metals have not yet been elucidated
Conventional wild duckweed cannot be directly applied to mechanism research, and it is difficult to carry out research on the ecological restoration of heavy metal-polluted water environment and the application of plant resistance to heavy metals. A kind of duckweed that can be used for mechanism research needs further research and development
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[0023] This invention was funded by the National Key Research and Development Program (2018YFD0901301), the National Natural Science Foundation of China Youth Project (31700443), and the Tianjin Fishery Industry Technology System Innovation Team (ITTFRS2017007).
[0024] The invention is funded by the National Natural Science Foundation of China (32071620) and the Natural Science Foundation of Tianjin (S20QNK618).
[0025] In the following technical solutions, the sources of the vectors pMD19-T and pCAMBIA are respectively stored in the Key Laboratory of Animal and Plant Resistance of Tianjin Normal University.
[0026] A kind of duckweed expression vector of neurotransmitter gamma-aminobutyric acid fluorescent probe iGABASnFR, its steps are as follows:
[0027] Step 1: Query and obtain the gene sequence of neurotransmitter γ-aminobutyric acid fluorescent probe (iGABASnFR)
[0028] The iGABASnFR gene sequence was obtained by querying on GenBank. The GenBank number is MH392466...
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The invention discloses a duckweed expression vector of a neurotransmitter gamma-aminobutyric acid fluorescent probe iGABASnFR and a construction method of the duckweed expression vector. The method comprises the following steps: designing an iGABASnFR gene specific primer by taking an iGABASnFR sequence synthesized from a whole gene as a template, connecting a product with a 35S promoter and an NOS terminator of the tobacco mosaic virus through PCR (Polymerase Chain Reaction) and enzyme digestion, and finally constructing the product onto a plant binary expression vector 1301 through double enzyme digestion. The construction of the duckweed expression neurotransmitter gamma-aminobutyric acid fluorescent probe iGABASnFR vector is finally determined and completed through PCR (Polymerase Chain Reaction) identification, double enzyme digestion identification and gene sequencing. The construction method disclosed by the invention has the characteristics of being simple, convenient and easy to operate, storing the original GUS gene of the 1301 vector, being easy to screen and the like, can be applied to the application research of the duckweed expression neurotransmitter gamma-aminobutyric acid fluorescent probe iGABASnFR, and is an indispensable key operation for the research of the adversity stress resistance mechanism of the duckweed.
Description
technical field [0001] The invention belongs to the technical field of bioengineering, and specifically relates to a duckweed expression vector of neurotransmitter gamma-aminobutyric acid fluorescent probe iGABASnFR and a construction method thereof. Background technique [0002] Duckweed is a monocotyledon water surface floating plant, which belongs to the Duckweed family and is widely distributed all over the world. It has the ability to enrich heavy metal cadmium ions. However, the physiological and molecular mechanisms of duckweed enriching heavy metals have not yet been elucidated. Conventional wild duckweed cannot be directly applied to mechanism research, and it is difficult to carry out research on the ecological restoration of heavy metal-polluted water environment and the application of plant resistance to heavy metals. A duckweed that can be used for mechanism research needs further research and development. Gamma-aminobutyric acid (abbreviated as GABA) is an im...
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IPC IPC(8): C12N15/82C12N15/66C12N15/65
CPCC12N15/8212C12N15/8251
Inventor 王珅杨琳马旭孙金生
Owner TIANJIN NORMAL UNIVERSITY