Marine microalgae chloroplast expression vector and application thereof

A technology of chloroplast expression and marine microalgae, which is applied in the field of marine microalgae chloroplast expression vectors, can solve the problems of high cost and complicated operation

Inactive Publication Date: 2011-01-19
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Chlamydomonas reinhardtii is the only eukaryotic algae that has successfully transformed chloroplasts. In this transformation system, the promoter and terminator derived from chloroplasts are used to regulate the expression of foreign genes, and the vector is introduced into the chloroplast genome by gene gu

Method used

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  • Marine microalgae chloroplast expression vector and application thereof
  • Marine microalgae chloroplast expression vector and application thereof
  • Marine microalgae chloroplast expression vector and application thereof

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Embodiment 1

[0093] (1) Design primers:

[0094] The upstream and downstream primers of homologous recombination sequence I are P1 and P2, respectively

[0095] P1: 5'-CGAGCTCACTGGGCGTAAAGCGTCTGT-3'

[0096] SacI

[0097] P2: 5'-GGGGTACCTTGGGCCATTCTGGATTTG-3'

[0098] KpnI

[0099] The upstream and downstream primers of homologous recombination sequence II are P3 and P4, respectively

[0100] P3: 5'

[0101] -ACGCGTCGACAACTGCAGCGGGGGTATAGCTCAGTTGG-3'

[0102] SalI

[0103] P4: 5'

[0104] -AACTGCAGACATGCATGCTTTCGTTACTCAAGCCGACATT-3'

[0105] PstI

[0106] (2) PCR amplification: using Phaeodactylum tricornutum genomic DNA as a template, wherein Phaeodactylum tricornutum (Phaeodactylum tricornutum) was purchased from Wuhan Institute of Hydrobiology, and the extraction process of Phaeodactylum tricornutum genomic DNA followed the UniversalGenomicDNAExtractionKitVer.3.0 reagent of Dalian Bao Biological Company box instruction manual.

[0107] The reaction system of homologous recomb...

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Abstract

The invention discloses a marine microalgae chloroplast expression vector and application thereof. The marine microalgae chloroplast expression vector contains a homologous recombinant sequence I, a target gene expression case and a homologous recombinant sequence II, wherein the target gene expression case is positioned between the homologous recombinant sequence I and the homologous recombinant sequence II; 1st to 972nd bp of the homologous recombinant sequence I are 3' part of rns, and 973rd to 1,097th bp are 5' part of trnI; and 1st to 196th bp of the homologous recombinant sequence II are 3' part of trnA, and 197th to 1,483rd bp are 5' part of rnl. The homologous recombinant sequence ensures that the expression vector can be induced into the marine microalgae chloroplast genome by an electric shock method, which is advantageous to the way of inducing the vector into the chloroplast genome, greatly reduces conversion cost, and provides technical support for realizing that marine microalgae chloroplast is used as a bioreactor to produce high value-added products.

Description

technical field [0001] The invention relates to the field of microalgae genetic engineering, in particular to a marine microalgae chloroplast expression vector and application thereof. Background technique [0002] Since the first transgenic plant tobacco came out in 1984, the transformation of exogenous genes into the plant nucleus has been the main research direction of plant genetic engineering. However, due to the low expression level of exogenous genes in the plant nucleus, the unstable expression in offspring, and the ecological security caused by the easy diffusion of nuclear genes with pollen, there are other problems. Compared with traditional nuclear transgenesis, chloroplast transformation has many advantages: there are a large number of chloroplast copies in each plant cell, which makes the overexpression of foreign genes difficult to achieve in nuclear gene transformation; foreign genes are targeted by homologous recombination After integration in the chloropla...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N1/13C12R1/89
Inventor 李宏业朱聪聪杨维东刘洁生
Owner JINAN UNIVERSITY
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