Artemisia apiacea4-(5'-cytidine diphosphate)-2-C-methyl-D-erythritol synthase coding sequence
A technology of cytidine diphosphate and coding sequence, which is applied in the field of coding sequence and coding sequence of Artemisia annua 4--2-C-methyl-D-erythritol synthase, and can solve problems such as no coding sequence reports yet, Achieve the effect of realizing large-scale production and increasing content
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[0023] Step 1, cloning of Artemisia annua 4-(5'-cytidine diphosphate)-2-C-methyl-D-erythritol synthase coding sequence (CMS)
[0024] (1) RNA extraction (RNA extraction)
[0025] Take the leaf tissue of Artemisia annua, grind it in liquid nitrogen, add it to a 1.5mL Eppendorf (EP) centrifuge tube filled with lysate, shake it fully, and extract total RNA according to the instructions of the TIANGEN kit. The quality of total RNA was identified by formaldehyde denaturing gel electrophoresis, and then the RNA content was determined on a spectrophotometer.
[0026] (2) Full-length cloning of the gene (Cloning of Full-length cDNA)
[0027] According to the conserved sequence of amino acids encoded by related genes in Arabidopsis, using the principle of homologous gene cloning, the RACE (Rapid Amplification of cDNA Ends) method (SMART of Clontech Company) was adopted. TM RACE kit) for full-length cDNA cloning in three stages:
[0028] ①Synthesis of first-strand cDNA
[0029] Take...
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