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Hansenula Polymorpha HU-11

产品信息
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    Description
    Using homology-mediated homologous integration to build a whey acid glycosides 5-phosphate
    decarboxylase gene (HURA3) destroyed recombinant H. polymorpha yeast strains HU-11. The
    strains used at home and abroad compared by the auxotrophic host strain mutagenesis, with high
    genetic stability, low reverse mutation characteristics; facilitate the conversion of genetic
    screening and recombinant strains, and keep the wild-type strain physiological and biochemical
    characteristics, is conducive to the recombinant strain culture and efficient expression of foreign
    proteins, with high value industrial applications. Hansenula gene is disrupted host bacteria strain
    HU11 URA3 gene DNA sequencing results showed: gene is disrupted resulting after the first 31
    bp insertion GAAGT five bases. GAAGT five nucleotide insertion produce frameshift
    mutations. Frameshift mutation resulted in all 254 codon 11 after being replaced.GAAGT five
    bases while producing reverse mutation probability is extremely small, experimental testing also
    demonstrated host bacteria strain HU11 reverse mutation rate is zero; this "square" Reply
    mutation rate of the host strain on the conversion filter is particularly advantageous.     Application of
    the above-mentioned gene knockout technology to build the URA3- auxotrophic host cell strain
    HU-11.
    Medium YM agar or YM broth
    YM Medium (Agar or Broth) Agar Medium YM Agar (BD
    271210)………………………..41 g DI
    Water………………………………….……1000 ml Autoclave at 121ºC. Broth
    Medium YM Broth (BD 271120)…………..………….21.0 g DI
    Water…………………………………..….1000 ml Autoclave at 121ºC. *This
    product is commercially available from BD. To make medium from scratch,
    follow formulation below: Yeast Extract………………………………….3.0 g Malt
    Extract…………………………………....3.0 g
    Dextrose………………………………………10.0g
    Peptone……………………………………….5.0 g Agar (if
    required)……………………………..20.0 g DI
    Water……………………………………….1000 ml pH 6.2 +/- 0.2
    Growth
    Conditions    		 Temperature: 24°C
    Atmosphere: Typical aerobic

    Recovery
    1. Obtain an YM agar plate with the appropriate antibiotic.
    2. Using a sterile pipette tip, touch the bacteria growing within the
    punctured area of the stab culture. (A sterilized wire loop or sterile
    toothpick can be used in place of a sterile pipette tip.)
    3. Run this tip lightly over a section of the plate, as shown in the figure,
    to create streak #1.
    4. Using another sterile pipette tip, pass through streak #1 and spread
    the bacteria over a second section of the plate, to create streak #2.
    5. Using a third sterile pipette tip, pass through streak #2 and spread
    the bacteria over the last section of the plate, to create streak #3.
    6. Grow overnight in a 24     o C incubator (unless a different growth temperature is indicated on the
    plasmid datasheet).
    7. In the morning, single colonies should be visible. If the bacterial growth is too dense, re-streak
    onto a new agar plate to obtain single colonies
    温馨提示：不可用于临床ZL。
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