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Functional protein POX01167, encoding gene thereof and application of functional protein POX01167 and encoding gene

A gene and encoding technology, applied in the field of functional protein POX01167 and its encoding gene and application, can solve the problem that the production of fungal cellulase cannot meet the needs of large-scale biorefining of lignocellulose

Active Publication Date: 2017-05-31
GUANGXI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the production of fungal cellulase is still far from meeting the needs of industrial-scale biorefining of lignocellulose.

Method used

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  • Functional protein POX01167, encoding gene thereof and application of functional protein POX01167 and encoding gene
  • Functional protein POX01167, encoding gene thereof and application of functional protein POX01167 and encoding gene
  • Functional protein POX01167, encoding gene thereof and application of functional protein POX01167 and encoding gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] Example 1. Discovery of functional protein POX01167 and its coding gene

[0102] After extensive sequence analysis, expression analysis and functional verification of the genome of Penicillium oxalicum (Penicillium oxalicum) strain HP7-1, a new protein and its coding gene were discovered. The literature on the genome sequence analysis of Penicillium oxalicum strain HP7-1 is as follows: Zhao, S., Yan, YS., He, QP., Yang, L., Yin, X., Li, CX., Mao, LC., Liao, LS., Huang, JQ., Xie SB., Nong, QD., Zhang, Z., Jing, L., Xiong, YR., Duan, CJ., Liu, JL., Feng, JX. Comparative genomics , transcriptomic and secretomic profiling of Penicillium oxalicum HP7-1 and its cellulase and xylanase hyper-producing mutant EU2106, and identification of two novel regulatory genes of cellulase and xylanase gene expression[J].Biotechnol Biofuel 2016,9:203; 1 The name in the literature is "Penicilliumoxalicum strain HP7-1". The preservation number of Penicillium oxalicum strain HP7-1 is: CGMCC ...

Embodiment 2

[0104] Example 2, Construction of POX01167 Gene Knockout Cassette

[0105] 1. Genomic DNA of Penicillium oxalicum strain HP7-1 was extracted.

[0106] 2. Using the genomic DNA obtained in step 1 as a template, a primer pair consisting of primer POX01167-left-F and primer POX01167-left-R was used for PCR amplification to obtain a 2299bp fragment upstream of the ORF of the POX01167 gene (see figure 1 Lane 1) for POX01167 left arm.

[0107] POX01167-left-F: 5'-GATCTGGTAGCCAATCCATTTC-3'; POX01167-left-R: 5'-TTTAGAGGTAATCCTTCTTTCTAGAGGCCGACGACTTGAGAAACTAT-3'.

[0108] 3. Using the genomic DNA obtained in step 1 as a template, a primer pair consisting of primer POX01167-right-F and primer POX01167-right-R was used for PCR amplification to obtain a 2336bp fragment downstream of the ORF of the POX01167 gene (see figure 1 Lane 3) for POX01167 right arm.

[0109] POX01167-right-F: 5'-TCCTTCAATATCATTCTTCTGTCGACGGCAGCCCGCGGAGTG-3';

[0110] POX01167-right-R: 5'-GACACGAAAAGGGGGGAAAAA...

Embodiment 3

[0122] Example 3, Construction and Verification of Penicillium oxalicum POX01167 Gene Deletion Mutant ΔPOX01167

[0123] 1. Construction of Penicillium oxalicum mutant strain ΔPOX01167

[0124] 1. Using Penicillium oxalicum strain HP7-1 as the starting bacterium, knock out its Ku70 gene to obtain the Penicillium oxalicum mutant ΔPoxKu70.

[0125] The documents that record the Penicillium oxalicum mutant ΔPoxKu70 and the specific steps of knocking out the PoxKu70 gene are as follows: Zhao, S., Yan, YS., He, QP., Yang, L., Yin, X., Li, CX., Mao, LC., Liao, LS., Huang, JQ., Xie, SB., Nong, QD., Zhang, Z., Jing, L., Xiong, YR., Duan, CJ., Liu, JL., Feng, JX. Comparativegenomic, transcriptomic and secretomic profiling of Penicillium oxalicum HP7-1 and its cellulase and xylanase hyper-producing mutant EU2106. and identification of two novel regulatory genes of cellulase and xylanase gene expression[J].Biotechnol Biofuel 2016, 9: The mutant ΔPoxKu70 is named "Penicilliumoxalicum mu...

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Abstract

The invention discloses functional protein POX01167, an encoding gene thereof and application of the functional protein POX01167 and the encoding gene. The protein provided by the invention is obtained from penicillium oxalicum and is named as POX01167 protein. The protein provided by the invention is as the following (a1) or (a2): (a1) protein consisting of an amino acid sequence as shown in a sequence 1 in a sequence list; and (2) protein formed by performing substitution and / or deletion and / or addition of one or more amino acid residues on the amino acid sequence in the sequence 1, having the same function and derived from the sequence 1. A gene for encoding the POX01167 protein (POX01167 gene) also belongs to the protection range of the invention. Experiments prove that the functional protein POX01167 plays a critical role in regulating and controlling the expression process of a cellulase gene and a xylanase gene of the penicillium oxalicum, and has application potentiality of increasing the yield of the cellulase and the xylanase.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a functional protein POX01167 and its coding gene and application. Background technique [0002] Lignocellulose is the most abundant renewable resource in the world. The use of lignocellulosic biorefinery to produce bioliquid fuels and biobased chemicals is of great strategic significance to alleviate today's energy crisis and deteriorating environmental problems. (Vishnu, M., Mala, R. Trends in bioconversion of lignocellulose: biofuels, platform chemicals & biorefinery concept [J]. Progress in Energy and Combustion Science2012, (38): 522-550; Miguel, V., Jose, L.G., Joaquina, L., Juna-Lius, R. Biofuels 2020: Biorefineries based on lignocellular materials [J]. Microb Biotechnol 2016, 9(5): 585-594). However, at present, the biorefinery using lignocellulose as raw material has not yet been commercialized, and the main bottleneck is the low yield and high cost of microbial cell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/385C12N15/31C12N15/80C12N1/15C12N9/24C12N9/42C12R1/80
CPCC07K14/385C12N9/2402C12N9/2437C12N9/248C12Y302/01004
Inventor 冯家勋闫语丝赵帅
Owner GUANGXI UNIV
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