Synthetase gene for T.fusca mycose and method for preparing mycose
A trehalose synthase, trehalose technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve problems such as DNA sequences and amino acid sequences are very different, enzyme activities are different, and bacterial species sources are different.
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Embodiment 1
[0110] 1. Cloning of trehalose synthase gene (treS)
[0111] Inoculate Thermobifida fusca into the following liquid medium (g / L): glucose 4.0, yeast extract 4.0, malt extract 10.0, pH 7.2, and then shake at a constant temperature of 50°C Shake on the bed for 24 hours, then centrifuge at 5000rpm for 10 minutes to collect the bacteria. Total DNA extraction from Thermobista spp. was then performed as described in "Molecular Cloning: A Laboratory Manual (Second Edition)" (Sambrook, et al. 1989, Molecular Cloning: a Laboratory Manual).
[0112] Design the following primers:
[0113] 1. Upstream primer (Sense primer):
[0114] 5-GAG CCATGG CCCAGGCACACCCGGAGTGGT-3
[0115] 2. Antisense Primer
[0116] 5-TGGTCT CTGCAG TCAATTCAACCGCAGCCAGTAATAG-3
[0117] '
[0118] 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.
[0119] The PCR ...
Embodiment 2
[0132] Using cassava starch as the starting material, after liquefaction by α-amylase, the maltose generated by 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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