Theanine synthetase gene and method for preparing theanine
The theanine synthase and the theanine technology are applied in the field of theanine synthase and the preparation of theanine by using the theanine synthase, which can solve the problems of high cost, cumbersome operation steps, large space occupation and the like, and achieve less side reactions, The effect of simple product purification steps
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[0041] 1.1 Test materials
[0042] The early fresh leaves of the tea tree variety Shucha were collected from the teaching and scientific research base of Anhui Agricultural University. After the samples were collected, they were quickly frozen in liquid nitrogen and then stored in a -80°C refrigerator for later use.
[0043] 1.2 Total RNA extraction and cDNA synthesis
[0044] The extraction of total RNA was carried out according to the CTAB lysis method of Shi Chengying et al. and purified using the Tiangen Plant RNA Purification Kit. Using the extracted tea tree total RNA as a template and Oligo(dT)18 as a primer, the first strand of cDNA was synthesized according to the instruction manual of PrimeScriptII RTase (Takara) reverse transcriptase.
[0045] 1.3 Gene cloning, prokaryotic expression and enzyme activity assay
[0046] (1) Gene source and GS1-1ORF verification
[0047] The gene sequence is derived from the EST screened from the radicle root library constructed by ...
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Abstract
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