Radioactivity resistant anomalous coccus trehalose synthase gene and trehalose making method
A technology of trehalose synthase and Deinococcus, which is applied in the direction of isomerase, genetic engineering, plant gene improvement, etc., can solve problems such as unapplied, different enzyme activities, and great differences in DNA sequence and amino acid sequence
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Embodiment 1
[0103] 1. Cloning of trehalose synthase gene (treS)
[0104] Deinococcus radiodurans was inoculated in the following liquid medium (g / L): glucose 5, yeast extract 5, peptone 10, sodium chloride (NaCl) 5, pH 7.4, Then shake on a constant temperature shaker at 30° C. for 24 hours, and centrifuge at 5000 rpm for 10 minutes to collect bacteria. Total DNA extraction of Deinococcus radiodurans was then performed as described in "Molecular Cloning: A Laboratory Manual (Second Edition)" (Sambrook, et al. 1989, Molecular Cloning: a Laboratory Manual).
[0105] Design the following primers:
[0106] 1. Upstream primer (Sense primer):
[0107] 5-GAG CCATGG CCCAGGCACACCCGGAGTGGT-3
[0108] 2. Antisense Primer
[0109] 5-TGGTCT CTGCAG TCAATTCAACCGCAGCCAGTAATAG-3
[0110] Then proceed according to the steps described in PCR Cloning Protocols: 94°C for 60 seconds, and then perform 27 ring diameters: 95°C for 45 seconds, 55°C for 60 seconds, and 72°C for 10 minutes.
[0111] The PCR...
Embodiment 2
[0125] Using cassava starch as the starting material, after liquefaction by α-amylase, the maltose generated by the action of β-amylase is reacted according to Example 1. The experiment shows that 100 kg of starch can obtain about 40 kg of trehalose. The conversion efficiency of moisture and impurities in starch is the same as that of using maltose directly as a substrate.
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