A kind of locust uridine diphosphate glucuronosyltransferase gene and its application
A uridine diphosphate glucose, transferase technology, applied in transferase, application, genetic engineering and other directions, can solve problems such as unfavorable agricultural environment, green, sustainable development, etc., and achieve the effect of reducing selectivity and fitness
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Embodiment 1
[0025] 2) The homogenate was transferred to a 1.5mL centrifuge tube, left standing at room temperature for 5min, and centrifuged at 13000r for 5min at 4°C.
[0026] 3) Transfer the supernatant to a clean 1.5mL centrifuge tube, add 200 μL of chloroform, and vortex for 15s. room temperature
[0029] 6) 4 ℃, 13000r centrifugal 2min, discard the filtrate.
[0034] Using PrimeScript
[0036] 2) After being incubated at 65°C for 5 min, rapidly cooled on ice.
[0038] 4) Mix slowly.
[0040] 6) Incubate at 95°C for 5min to inactivate the enzyme, place on ice, and store the cDNA at -20°C.
[0049] PCR reaction conditions are as follows: 95°C for 3 min; 95°C for 30s, 55°C for 30s, 72°C for 1.5min, 35 cycles; 72°C
[0059] The sequencing results show that: PCR amplification obtains a DNA fragment with a size of 1561 bp.
Embodiment 2
[0064] Primers were designed according to the cloned gene fragments, and a T7 promoter was introduced at the 5' end of the primers. The primer sequences are as follows:
[0070] PCR reaction conditions are as follows: 95°C for 3min; 95°C for 30s, 55°C for 30s, 72°C for 1min, 35 cycles; 72°C for 10min;
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