33results about How to "Genetic manipulation is simple" patented technology

The invention provides EV71virus-like particles as well as preparation method and application of the EV71virus-like particles. The method comprises the following steps of: connecting a P1 protein gene and 3CD protease gene of an EV71 virus with predicated mean vote (PMV) plasmid to build PMV-P1-3CD recombinant expression plasmid; turning into hansenula polymorpha AU-0501 expression bacterial strain to obtain an AU-PMV-P1-3CD recombinant expression bacterial strain; fermenting and culturing the recombinant expression bacteria strain and performing inducible expression on EV71 virus-like particle proteins by using methanol; centrifugally gathering thallus to perform homogenate crushing at high pressure; and purifying the supernate through ion-exchange column chromatography, hydrophobic chromatography and sieve chromatography to obtain the EV71virus-like particles. EV71virus-like particle vaccines provided by the invention has high immunogenicity, security, immunological characteristics and biological activity; and the EV71virus-like particles can be prepared in a large scale and purified and used for preparing vaccines for preventing EV71 infection, and has good economic value and application prospect.

The present invention provides EV71 virus-like particles and a preparation method and application thereof. The method comprises: connecting a P1 protein gene and a 3CD protease gene of an EV71 virus with a PMV plasmid to construct a PMV-P1-3CD recombinant expression plasmid; then transforming a Hansenula polymorpha AU-0501 expression strain with the PMV-P1-3CD recombinant expression plasmid to obtain an AU-PMV-P1-3CD recombinant expression strain; fermenting and culturing the recombinant expression strain, and inducing the recombinant expression strain to express the EV71 virus-like particle protein with methanol; centrifuging and collecting mycelia for homogeneous breakage at a high pressure; and purifying the supernatant through ion-exchange chromatography, hydrophobic chromatography, and molecular sieve chromatography, so as to obtain EV71 virus-like particles.

The invention relates to high-temperature riboflavin-producing engineering bacteria and its construction method and application. The engineering bacterium is Escherichia coli or Geobacillus, and carries an expression vector expressing Geobacillus riboflavin synthesis gene cluster. Riboflavin can be produced by high-temperature riboflavin-producing Escherichia coli engineering bacteria, and the riboflavin production in the fermentation broth can reach up to about 276mg / L. Up to about 50mg / L. The engineering bacterium provided by the invention has a shorter production cycle than the existing riboflavin engineering bacterium, saves energy, can be used for riboflavin production at low cost, and provides valuable strain resources for producing riboflavin under high temperature conditions.

The invention discloses a ship branch pipeline path planning method. The method comprises the steps of on the basis of simplifying a device ad a pipeline model, establishing a network graph between two points referring to a visible graph method in mobile robot path planning; setting branch pipelines containing N connection points, establishing the network graph between every two points of the N connection points, and combining update data information, thereby obtaining the total network graphs among the N points; establishing a population based on a Steiner point genetic algorithm, setting genetic algorithm parameters and then starting iteration to carry out optimization; determining the equivalent lengths of the pipelines and endowing different weight coefficients to different pipelines; evaluating the fitness value of each chromosome by use of a distance heuristic algorithm; judging whether set irritation times arrives or not; outputting optimum paths, finishing path planning, updating pipeline code information in a storage document, and realizing three-dimensional visualization of the ship branch pipelines through combination of three-dimensional design software. The method has very high search efficiency and can satisfy the practical demands of ship branch pipeline path planning better. The layout problem of the branch pipelines is solved.

The invention discloses a cloud workflow scheduling optimization method based on a hierarchical and load balancing genetic algorithm, comprising the following steps: obtaining information required for scheduling optimization; calculating the hierarchical value of a task; initializing a contemporary population; Population, perform mutation operation on the new population, decode and calculate the fitness value and improve it by means of hierarchy and load balancing, select N different individuals from the contemporary and new populations to form a new contemporary population, until the evolution termination condition is met; Output scheduling optimization scheme. On the premise of realizing global search, the invention adopts serial individual decoding based on insertion mode, individual improvement based on hierarchy and load balancing, parameterized uniform crossover based on preference and other methods to improve the optimization ability and search efficiency of the algorithm.

The invention belongs to the technical field of molecular biology and biological chemistry and in particular relates to preparation of recombinant protein of crassostrea gigas cell wall hydrolase and application of the prepared recombinant protein. The recombinant protein of the crassostrea gigas cell wall hydrolase is prepared by utilizing a yeast expression system. The recombinant protein of the crassostrea gigas cell wall hydrolase, which is prepared by utilizing a yeast expression system, has a remarkable inhibition effect on the growth of escherichia coli, aeromonas hydrophila, staphylococcus aureus, vibro splendidus, vibrio vulnificus, vibrio alginolyticus, pseudomonas putida and pseudomonas aeruginosa. The recombinant protein provided by the invention can be used for developing bacteriostatic agents.

The invention relates to a genetically engineered bacterium with a high production of farnesene, a construction method and application thereof, and belongs to the technical field of microorganisms. In order to solve the problems of low catalytic efficiency of heterologous MVA downstream pathway and low farnesene production in the process of synthesizing farnesene in Escherichia coli, the present invention constructs plasmids pACYC-mvaE-mvaS-ispA-AaFS, pTrcLower-ΔIDI, pTrcLower- AaIDI and pET28a-AaFS-ispA-SlIDI / AaIDI (IDI genes are from tomato and Artemisia annua respectively), the above plasmids were combined and transformed into Escherichia coli, and the culture conditions were improved, and the prepared genetically engineered bacteria could significantly improve the synthetic yield of farnesene , which is conducive to promoting the industrialization process of biosynthesis of farnesene.