32 results about "Yeast chromosome" patented technology

The invention discloses a yeast recombinant human type I collagen alpha 1 chain protein, a synthesis method and application thereof. The recombinant human type I collagen alpha 1 chain protein of theinvention consists of an N-terminal affinity purification marker, a human type I collagen alpha 1 chain mature peptide sequence and a C-terminal affinity purification marker. The bispecific affinity purification markers designed at the two terminals are beneficial to purification of the recombinant protein and are also beneficial to detection and full-length identification of the recombinant protein. The synthesis method comprises the following steps: firstly optimizing collagen gene sequence by utilizing common codons of pichia pastoris at gene level and synthesizing the whole gene artificially, then constructing a recombinant vector through PCR, enzyme digestion, connection and the like, and integrating into yeast artificial chromosome to construct recombinant pichia pastoris engineeringbacteria for secretory expression of the recombinant human type I collagen alpha 1 chain protein. The bispecific affinity purification marker carried by the recombinant human type I collagen alpha 1chain protein of the invention can be subjected to two-step specific purification to easily obtain high-purity products.

The invention discloses design and construction methods and application of standardized bio-elements. The construction method of the standardized bio-elements comprises the following steps that 1, a target genophore group, a promoter carrier group and a terminator carrier group are constructed. A series of standardized bio-elements are designed and constructed based on a yeast system, restriction enzymes II can be utilized for assembling the standardized bio-elements into an ultralong foreign DNA fragment, the foreign DNA fragment is a target product synthesis route composed of multiple genes, and the fragment is integrated to a specific site of a yeast chromosome through yeast homologous recombination for stable expression; a heterogenetic source in the target product yeast can be synthesized, optimum expression of the target product can be achieved by optimizing the promoters of all the genes, and therefore unique advantages and great significance are achieved in the optimization process of target product synthesis.

The invention relates to the field of microorganisms, in particular to a transfer method for saccharomyces cerevisiae chromosomes. A synthetic No.V chromosome saccharomyces cerevisiae strain and a synthetic No.X saccharomyces cerevisiae strain are fused into a diploid strain through an endoreduplication backcross method, the wild No.V chromosome and No.X chromosome are induced to be lost through galactose, and screening is conducted through an SC+5-FOA culture medium; a saccharomyces cerevisiae haploid strain carrying the synthetic No.V and No.X chromosomes is obtained through spore splitting. Compared with existing achievements, the transfer method has the advantages that transfer of the saccharomyces cerevisiae chromosomes is rapidly achieved; cross swap of sister chromatids is avoided, and the saccharomyces cerevisiae haploid strain with the chromosomes transferred is obtained.

The invention relates to the field of genome engineering and discloses a method for repairing the anomaly of a yeast chromosome structure. According to the method disclosed by the invention, yeast chromosomes, which have chain exchange at a design region, are rapidly screened by utilizing an efficient homologous recombination mechanism and a screening label of yeast, and exchange between an sequence with the abnormal yeast chromosome structure and a normal chromosome sequence is realized, so that the aim of efficiently repairing the variation of the yeast chromosome structure in a large scale is realized.

The method for yeast cell to express human interleukin 10 (hIL-10) has the technological scheme including artificially synthesizing all-gene sequence of hIL-10 including signal-carrying hIL-10 sequence, named shIL-10, and no-signal hIL-10 sequence, named nshIL-10; adding to the ends with EcorR I enzyme incising sequence and Not I enzyme incising sequence or with EcorR I enzyme incising sequences; double incising with EcorR I and Not I the integral expression vector pPIC3.5K and Ppic9K of destination gene shIL-10, nshIL-10 and Pichia yeast; constituting recombinant plasmid pPIC3.5K / shIL-10 and Ppic9K / nshIL-10; converting Pichia yeast cell via electrical perforation and plasmodic granule process; integrating recombinant plasmid and destination gene to yeast chromosome; and applying methanol to induce Pichia yeast for intracellular soluble expression and extracellular secretion type expression. The present invention is superior to expression in colibacillus.

The invention belongs to the technical field of molecular biology, in particular to a method for producing genetic engineering recombinant 192 peptide human growth hormone. The technical scheme is as follows: (1) Obtaining the 192-peptide human growth hormone gene: using the pHil-D2 plasmid as a shuttle plasmid, the growth hormone cDNA is transferred into yeast cells, and through homologous recombination with genes on the chromosomes of yeast cells, The genetically engineered hGH yeast was constructed; (2) expression system: the strain of Pichia pastoris was used as the expression host to express 192 peptide human growth hormone in the way of intracellular expression; (3) the hGH yeast was induced to express and produce human growth hormone.

The invention relates to the technical field of biology, and specifically discloses a method for knocking down brewing yeast chromosome. The method is characterized in that the complete yeast chromosome is cut according to the CRISPR / Cas9 base technology; a specificity target close to centromere is selected; a corresponding guide RNA is designed to guide Cas9 protein to generate an incision closeto the centromere, thus realizing the knocking down of the whole chromosome. Compared with the conventional method for losing the whole chromosome in manners such as inducing through a Ga1 promoter, and reductional division, the method has the advantages that the brewing yeast chromosome is simply, efficiently and quickly cut; the cross exchange of sister chromatids can be avoided; the chromosome-knocked-down homozygous brewing yeast diploid strain can be obtained.

The invention relates to an efficient yeast chromosome fusion method. The invention discloses a method for performing yeast chromosome fusion by using a CRISPR / Cas9 system and a homologous recombination system in yeast. The method of the invention can realize rapid and efficient chromosome fusion.

The invention relates to the technical field of biology and in particular discloses a method for knocking out a saccharomyces cerevisiae chromosome. According to the method, a Vika / vox technology is utilized for removing a complete yeast chromosome, and chromosome centromeres are specifically recombined into a ring by virtue of loci, so that knockout of the whole chromosome is realized. Compared with a method for realizing elimination of the whole chromosome in a reduction mitosis manner, the method disclosed by the invention has the advantages that cutting of the saccharomyces cerevisiae chromosome is more simply, efficiently and rapidly realized, crossing over of sister chromatids is avoided, and a saccharomyces cerevisiae homozygous diploid bacterial strain with the chromosome knocked out is obtained.