Hansenula Polymorpha HU-11_详细资料

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订购或咨询热线 TEL：18801458894 QQ：3404177254
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
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