Hansenula Polymorpha HU-11 BioVector NTCC质粒载体菌种细胞基因保藏中心

Hansenula Polymorpha HU-11 BioVector NTCC质粒载体菌种细胞基因保藏中心 NTCC690054 Hansenula Polymorpha HU-11 1Stab culture, Storage:4℃


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.

BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心 www.biovector.net [Supplier来源] http://www.biovector.net