Metallothionein-like protein gene of Tamarix androssowii Litw.
A technology of metallothionein and tamarisk, applied in the field of genetic engineering, can solve the problems of lack of genes, strong stress resistance of salt-resistance-related genes, limited types and numbers of genes of stress-resistant plants, etc.
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[0022] By constructing a suppressive subtractive cDNA library, then randomly sequencing the library clones, and performing BlastX analysis on the sequencing results, the specific steps are:
[0023](1) Construction of a suppressive subtractive library. 2 μg of mRNA from the drought treatment and control materials were taken respectively, and 5'TTTTGTACAAGCTT30N 3' was used as a primer to synthesize the first and second strands of cDNA using the cDNA synthesis Kit (Bao Bioengineering (Dalian) Co., Ltd.). The synthesized double-stranded cDNA was digested with RsaI, and the digested products were purified and precipitated, dissolved in 5.5 μL of ultrapure water, and 0.5 μL of each was used for agarose gel electrophoresis detection. The drought-treated cDNA was used as a tester, and the control cDNA was used as a driver. Take 1 μL of the tester cDNA and add 5 μL of water to dilute, take 2 μL of the diluted solution to connect with Adaptor1 and Adaptor2 respectively, the connectio...
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