CN110041417A

CN110041417A - A kind of hexose transport albumen and its encoding gene and application

Info

Publication number: CN110041417A
Application number: CN201910365074.8A
Authority: CN
Inventors: 成金桃; 文苏莹; 别之龙; 鲍天旸; 曾祥伟
Original assignee: Huazhong Agricultural University
Current assignee: Huazhong Agricultural University
Priority date: 2019-04-30
Filing date: 2019-04-30
Publication date: 2019-07-23

Abstract

The invention discloses a kind of albumen relevant to hexose transport, which has the amino acid sequence as shown in sequence table SEQ ID NO:2, and the invention also discloses the encoding gene of the albumen and applications.By clone's hexose transport protein gene CsHT11 sequence and carry out the heterologous functional verification of yeast, the experimental results showed that, CsHT11 albumen has the function of transporting glucose, fructose, galactolipin, mannose, and it is a linear process that Yeast transformants, which absorb glucose, the optimal pH for absorbing glucose is 4-6.These results illustrate that there is CsHT11 albumen very high hexose to absorb vigor, the substrate absorbed simultaneously is more extensive, it can be used for the genes of brewing yeast engineering improvement for the industry of making wine, CsHT11 may play a significant role in cucumber growth course simultaneously, can provide thinking and reference for cucumber glycometabolism and other melon crop sugar accumulation correlative studys.

Description

A kind of hexose transport albumen and its encoding gene and application

Technical field

The present invention relates to a kind of hexose transport albumen and its encoding gene and applications.

Background technique

(HTs) STPs belongs to MST (monosaccharide transport protein family) subtribe, has 12 transmembrane domains.In arabidopsis, AtSTP family represents the hexose transport albumen of most feature, and AtSTP family is located on plasma membrane, have promote hexose from Apoplastic spaces transport the function of cell.The type and activity that sugar is transported between different hexose transport protein family members have Biggish difference, the AtSTP2 in arabidopsis mainly transport hexose and pentose；AtSTP7 mainly transports L-arabinose and D- wood Sugar；AtSTP8 transports glucose, galactolipin, mannose；The albumen of AtSTP10 coding can make glucose, galactolipin, mannose For substrate；AtSTP12 transports glucose, galactolipin, mannose and xylose；AtSTP 14 transports glucose, fructose, galactolipin； AtSTP13 can transport glucose, fructose, galactolipin and mannose.VvHT5 transports glucose and fructose in grape； The high affine glucose transporter for relying on H+ of the coding of VvHT1,4,5.The affine transhipment glucose of LeHT1, LeHT2 high in tomato, Low affine transhipment fructose.CsHT3 transports glucose, galactolipin and mannose, CsHT4 and specifically transports galactolipin (Cheng in cucumber et al.2015a)。

A large amount of carbohydrate is needed to supply during pollen development and pollen tube rapid elongation, and nutrition supply is insufficient It may cause the abortion of pollen, and then influence Pollination Fertilization and solid.So hexose transport albumen has not in this process The effect that can or lack.In thaliana flower tube cell, fructose can be transported by AtSTP6, and glucose can not only pass through AtSTP6 can also be transported by AtSTP4, AtSTP8, AtSTP9, AtSTP10, AtSTP11.CsHTl is an energy in cucumber Dependent form hexose transport albumen is measured, it is main to transport glucose and galactolipin, specifically expressing (the Cheng et in pollen al.2015b).And specifically expressed hexose transport albumen may have higher hexose transport activity in pollen, it may be in flower It plays an important role in powder growth course.

Inventor has applied for the hair of one " albumen relevant to hexose transport and its encoding gene and application " in 2011 Bright patent (application number 201110307028.6), the hexose transport albumen are the CsHT1 from cucumber, which has Portugal The function of grape sugar transport can make the yeast mutants of hexose trafficking defect restore the absorption function of glucose.But inventor It is further discovered that CsHT1 albumen is poor to the transhipment effect of other hexoses such as fructose, galactolipin, in practical applications by certain Limitation.

Present invention finds on cucumber another in pollen and the specifically expressed hexose transport PROTEIN C sHT11 of pollen tube, Compared with CsHTl, the sugared substrate of CsHT11 Protein transport is more, and has very high hexose transport activity, which may be There is prior effect in pollen and pollen tube development, in addition, the albumen may have more in sugaring and brewing alcohol industry Good application potential.By being carried out to domestic and international patent document, periodicals and magazines and other documents published (such as internet) Retrieval, the present invention have not been reported at home and abroad, we, which also not publicly deliver, is related to the article of the content of present invention, and the present invention is at home The outer public is unknown.

Summary of the invention

The object of the present invention is to provide a kind of hexose transport albumen and its encoding genes.

Hexose transport albumen provided by the present invention, entitled CsHT11 (Hexose Transporter) derive from calabash Lu Ke, Cucumis, cucumber (Cucumis sativus L.) close thorn in kind Xintai City, are following protein 1) or 2):

1) protein that the amino acid sequence shown in SEQ ID NO:2 forms；

2) by the amino acid sequence of SEQ ID NO:2 by one or several amino acid residues substitution and/or missing and/ Or addition and the protein as derived from 1) relevant to hexose transport.

SEQ ID NO:2 is made of 515 amino acid residues.

Above-mentioned sequence can be artificial synthesized, can also first synthesize its encoding gene, then carries out biological expression and obtain.It is above-mentioned 2) in The encoding gene of CsHT11 can be by by the DNA sequence dna shown in the 5 ' end 1-1548 bit bases shown in SEQ ID NO:1 The codon of middle one or several amino acid residues of missing, and/or the missense mutation of one or several base-pairs of progress obtain.

It is above-mentioned to also belong to protection scope of the present invention with hexose transport GAP-associated protein GAP encoding gene.

With the encoding gene of hexose transport GAP-associated protein GAP concretely following 1) -3) in any gene:

1) its nucleotide sequence is sequence shown in SEQ ID NO:1；

2) it can hybridize under strict conditions with the SEQ ID NO:1 DNA sequence dna limited and encode above-mentioned and hexose transport phase Close the DNA molecular of albumen；

3) there is 90% or more homology with the gene 1) limited, and encodes above-mentioned and hexose transport GAP-associated protein GAP DNA molecular.

It is described 3) in gene, 1) or 2) 95% or more homology is preferably formed with the gene limited.

Sequence shown in SEQ ID NO:1 is made of 1548 base-pairs, is the open reading frame of CsHT11 gene (ORF), encoding amino acid sequence is CsHT11 albumen shown in SEQ ID NO:2.

The primer pair for expanding above-mentioned CsHT11 segment also belongs to protection scope of the present invention, draws as nucleotide sequence is following Object pair:

CsHT11F:5'-ATGGCTGGAGGAGGATTTGTTGCTGAAG-3'(SEQ ID NO:3)

CsHT11R:5'-TCAAGCATCGTCGTCATGCTTCCCC-3'(SEQ ID NO:4)

Containing above-mentioned with hexose transport related protein encoding gene and the recombinant vector of promoter, transgenic cell line and again Group bacterium also belongs to protection scope of the present invention.

The recombinant expression carrier of CsHT11 gene can be contained with existing plant expression vector construction.The plant expression carries Body includes double base agrobacterium vector and the carrier etc. that can be used for plant micropellet bombardment, as pCAMBIA3301, pCAMBIA1300, The derivative plant expression vector of pBI121, pBin19, pCAMBIA2301, pCAMBIA1301-UbiN, PDR196 or other.It carries There is the present invention can be by Ti-plasmids, Ri plasmid, plant virus with hexose transport related protein encoding gene CsHT11 expression vector The conventional biology methods such as carrier, directly delivered DNA, microinjection, conductance, mediated by agriculture bacillus are transformed into plant cell or tissue In.

When recombinant plant expression vector gene constructed using CsHT11, any one can be added before its transcription initiation nucleotide Enhanced, composing type, organizing specific type or inducible promoter are planted, such as cauliflower mosaic virus (CAMV) 35S promoter, general life Plain gene Ubiquitin promoter (pUbi) etc., they can be used alone or are used in combination with other plant promoters；May be used also The Marker positioned using the gene that CsHT11 is encoded as plasma membrane, is transferred in subcellular expression vector, using protoplast and The method of epidermal tobacco injection examines whether target gene is located in cytoplasma membrane.

For the ease of transgenic plant cells or plant are identified and screened, plant expression vector used can be carried out Processing can express gene (gus gene, fluorescein of the enzyme or luminophor that can produce color change as being added in plant Enzyme gene etc.), resistant antibiotic marker (gentamicin marker, kanamycins marker etc.) or it is anti-chemistry examination Agent marker gene (such as anti-herbicide gene).From the security consideration of genetically modified plants, any selected marker base can be not added Cause directly screens transformed plant with adverse circumstance.

The recombinant expression carrier be concretely inserted between the multiple cloning sites of pDR196 sequence 1 in sequence table from 5 ' The recombinant expression carrier pDR196-CsHT11 that the deoxynucleotide of end 1-1548 obtains.

Present invention discover that a CsHT11 albumen relevant to hexose transport, thin using the gene constructed Asia for encoding the albumen Born of the same parents positioning carrier CsHT11-pH7LIC5.0-ccdB rc-N-eGFP, is injected by epidermal tobacco, it is found that it is located in plasma membrane On.On the other hand building Yeast expression carrier pDR196-CsHT11, is transferred to the yeast mutants of hexose defect In EBY.VW4000, it is demonstrated experimentally that the yeast mutants for being transferred to pDR196-CsHT11 carrier can restore yeast glucose, Fructose, galactolipin, mannose absorption function, and the yeast mutants for being only transferred to PDR196 zero load cannot then restore yeast or more Sugar source absorption function illustrates that CsHT11 albumen has the function of transporting glucose, fructose, galactolipin, mannose.Further grind Studying carefully discovery pDR196-CsHT11-EBY.VW4000 Yeast transformants to absorb glucose is a linear process, while to difference The optimal pH that the experiment of pH Preference shows that CsHT11 absorbs glucose is 4-6.Expression and function to CsHT11 gene carry out deep Entering research helps to illustrate effect of the gene in Cucumber Pollen growth course, further mentions for research cucumber glycometabolism mechanism Theoretical foundation is provided for theoretical foundation, and for the work of cucumber high-efficient breeding.

Detailed description of the invention

Fig. 1 is the agarose gel electrophoresis figure after CsHT11 gene magnification.

Fig. 2 is pDR196-CsHT11-EBY.VW4000 Yeast transformants in glucose, fructose, galactolipin, mannose training Support the growing state on base.

Fig. 3 is for pDR196-CsHT11-EBY.VW4000 Yeast transformants with time change on dextrose culture-medium Growth curve.

Fig. 4 be pDR196-CsHT11-EBY.VW4000 Yeast transformants cultivated on dextrose culture-medium 8 hours after not With the growth curve under pH.

Fig. 5 is subcellular of the CsHT11-pH7LIC5.0-ccdB rc-N-eGFP under laser co-focusing in epidermal tobacco Positioning result.GFP shines for gene, and Bar=10 μm.

Specific embodiment

The present invention is further explained in the light of specific embodiments.Experimental method used in following embodiments is such as It is conventional method without specified otherwise.Material used in embodiment, reagent etc., unless otherwise specified, commercially It obtains.

The clone of embodiment 1, CsHT11 translated region overall length

The seed of the close thorn in cucumber variety Xintai City is seeded in equipped with turf, vermiculite and perlite (volume ratio 3:1:1) In mixed-matrix, in vinyl house, Routine Management takes definite value after plant blossom result when plant grows to four leaves wholeheartedly Root, stem, leaf, female flower, male flower, carpopodium, seven different tissues of fruit sample and extract total serum IgE respectively, reverse transcription cDNA.With Above-mentioned cDNA is template, and the CsHT11 reference sequences that are obtained according to Cucumber germplasm sequence alignment design PCR primer:

CsHT11F:5'-ATGGCTGGAGGAGGATTTGTTGCTGAAG-3'

CsHT11R:5'-TCAAGCATCGTCGTCATGCTTCCCC-3'

The code area overall length of hexose transport protein gene is cloned by the method for PCR.Reaction mixture is as follows: high fidelity enzyme 25 μ l of Mix, 2 μ l of upstream primer (10uM), 2 μ l of downstream primer (10uM), template (cDNA of reverse transcription) 2 μ l, ddH₂O 19μ l。

PCR reaction condition: first 94 DEG C of initial denaturation 5min, then 94 DEG C of denaturation 30s, 56 DEG C of renaturation 30s, 72 DEG C extend 2min, totally 35 recycle, last 72 DEG C of extensions 5min.

50 μ L PCR products are taken to carry out agarose gel electrophoresis, the results showed that only to expand 1548bp's or so in male flower Purpose band (Fig. 1).Target fragment is cut, is connected on pEasy-Blunt carrier and is sequenced after purification and recovery.Through homology The 1548bp segment that acquisition is primarily determined after comparative analysis is the full length coding region of CsHT11 gene.

By another pollen-specific expressing gene CsHT1 (103044533 A of CN) in the sequence and cucumber and intend south Male flower expresses special STP (HT) sequence and carries out homologous comparison analysis in mustard, as a result, it has been found that these gene orders are with higher Homology, wherein CsHT1 and CsHT11 has 72% homology.

The transport function verifying and analysis of embodiment 2, cucumber hexose transport PROTEIN C sHT11 in yeast

1, the building of recombinant expression carrier pDR196-CsHT11

1) expression vector and bacterial strain

Expression vector: pDR196

The yeast mutants bacterial strain of hexose absorption function missing: EBY.VW4000

2) design of primers

PDR196-CsHT11F:5'-TCCCCCGGGATGGCTGGAGGAGGATTTGTTGCTGAAG-3'(wave part For Sma I restriction enzyme site)

PDR196-CsHT11R:5'-CCGCTCGAGTCAAGCATCGTCGTCATGCTTCCCC-3'(wave part is Xho I restriction enzyme site)

3) PCR amplification: using pDR196-CsHT11F/R primer amplification, and the clone of overall length is specifically translated with example 1.

4) gene and carrier digestion

37 DEG C of overnight digestions.

5) gene is connect with carrier

16 DEG C of reaction 10h.

6) large intestine converts

(1) it prepares and contains carrier resistance-ammonia benzyl mycin solid LB media (2) step of converting

A) in superclean bench, take 50 μ L competent escherichia coli cells in the 1.5mL centrifuge tube of sterilizing, then plus 10 μ L connection products.It gently inhales and plays mixing, immediately ice bath 25min.

B) 42 DEG C of water-bath 90s, do not shake, later quick ice bath 2min.

C) 500 μ L LB liquid mediums (not added with antibiotic) are added in super-clean bench, mix.37 DEG C, the training of 200rpm shaking table 1h is supported, cell recovery is made.

D) in superclean bench, each plate applies about 100-150 μ L transformed bacteria, is uniformly coated with bacterium solution with spreader On solid LB plate.

E) plate for sealing mouth is inverted in 37 DEG C of incubators and is cultivated 10-13 hours.

7) bacterium colony PCR is identified

Picking monoclonal bacterial plaque accesses 37 DEG C of shaking table cultures in 800 μ L LB liquid mediums (containing antibiotic), muddy to shaking Bacterium solution carries out PCR identification.By the bacterium solution for identifying purpose band send out sequencing and the accurate bacterium solution of sequencing result expand it is numerous, It is spare for saving and extracting pDR196-CsHT11 recombinant plasmid.

2, pDR196-CsHT11 recombinant plasmid transformed mutant yeast strains EBY.VW4000

1) prepared by mutant yeast strains EBY.VW4000 competence

A) the well-grown EBY.VW4000 monoclonal yeast bacterial plaque of picking is in 1mL YPD fluid nutrient medium (2% albumen Peptone, 1% yeast extract, 2% maltose) in, 30 DEG C, 200rmp shakes bacterium until muddy.

B) above-mentioned bacterium solution is transferred to the fresh YPD fluid nutrient medium of 20mL, 30 DEG C, 200rmp shakes bacterium 12-15h, makes OD₆₀₀> 1.5。

C) it takes above-mentioned bacterium solution 2mL to be transferred in 40-50mL YPD fluid nutrient medium, makes OD₆₀₀Between 0.2-0.3,30 DEG C, 200rmp shakes bacterium 3-4h, makes OD₆₀₀Reach 0.6-0.8, at this time transformation efficiency highest.

D) bacterium solution is dispensed with the 50mL centrifuge tube of sterilizing, room temperature 1500g is centrifuged 10min, collects thallus, abandons supernatant.

E) cell is resuspended with the 1X TE Buffer (pH7.4) of 5mL sterilizing, room temperature 1500g is centrifuged 5min, collects cell and abandons Supernatant.

F) it is repeated once step e).

G) cell is resuspended with the 0.1M LiAC/TE Buffer of 1mL sterilizing and is transferred in the centrifuge tube of 2mL sterilizing, 30 DEG C Incubated cell 1h, room temperature 1500g are centrifuged 5min, collect cell and abandon supernatant.

H) cell is resuspended with the 0.1M LiAC/TE Buffer of 1mL sterilizing, room temperature 13000g is centrifuged 15s, collects cell and abandons Supernatant.

I) cell is resuspended with the 0.1M LiAC/TE Buffer of 500 μ L sterilizing, dispenses yeast cells, institute with 100 μ L/ pipes Obtaining cell is competent yeast cells.

2) competent yeast cells convert

A) the Yeast maker host DNA (SS-DNA) and 100ng Plasmid DNA to be transferred to of 5 μ L denaturation are drawn, mixing is equal It is even.

B) 100 μ L competent yeast cells are added, it is soft to mix.

C) 500 μ L 0.1M LiAC/PEG are added, it is soft to mix.

D) 30 DEG C of constant-temperature incubation 30min are placed in, are mixed by inversion every 10min.

E) 20 μ L DMSO are added, it is soft to mix.

F) 42 DEG C of water-bath 15min, are mixed by inversion every 5min.

G) 18000rpm is centrifuged 30s.

H) supernatant is abandoned, 1mL YPD culture medium is added, 18000rpm is centrifuged 30s.

I) supernatant is abandoned, 200 μ L 0.9%NaCl solution is added, thallus is resuspended.

J 100 μ L coated plates) are taken.

K) it is cultivated 2-3 days in 30 DEG C of insulating boxs.

3, pDR196-CsHT11-EBY.VW4000 Yeast transformants growth Drop test experiment

The eugonic positive pDR196-CsHT11-EBY.VW4000 yeast monoclonal of picking is in the SD- of 2%moltose In ura (pH6.0) fluid nutrient medium, 30 DEG C of cultures to OD overnight₆₀₀> 1.5 draws a small amount of bacterium solution fresh containing 2%Maltose SD-ura (pH6.0) culture medium in adjust concentration OD₆₀₀=0.2,30 DEG C are cultivated to OD₆₀₀=0.6, contain 2% with sterilizing Maltose SD-ura (pH6.0) culture solution dilutes 1,10,100,1000 times respectively, mixes well, and draws 5 μ l and puts respectively 2%maltose (maltose), 2%Glucose (glucose), 2%Frucose (fructose), 2%Galactose (galactolipin), 2%Mannose (mannose) is 2-4 days observation experiment knots of 30 DEG C of cultures on SD-ura (pH6.0) culture medium of sole carbon source Fruit, with pDR196 empty carrier recombination yeast pDR196-EBY.VW4000 as control.

As a result as shown in Figure 2, the results showed that using maltose as sole carbon source, Yeast transformants pDR196- CsHT11-EBY.VW4000 and pDR196 empty carrier recombination yeast pDR196-EBY.VW4000 can normal growth, but Respectively using glucose, fructose, galactolipin, mannose as sole carbon source in the case where, only Yeast transformants pDR196- CsHT11-EBY.VW4000 can be with normal growth, and pDR196 empty carrier recombination yeast pDR196-EBY.VW4000 cannot be just It is frequently grown, illustrates that being transferred to for CsHT11 gene has restored yeast mutants EBY.VW4000 to glucose, fructose, galactolipin and sweet Reveal the absorption function of sugar.

4, CsHT11 transports the linearly and pH value dependency analysis of glucose

By pDR196-EBY.VW4000 and pDR196-CsHT11-EBY.VW4000 yeast in SD-ura+2%maltose It is cultivated to OD for 30 DEG C in culture medium₆₀₀=0.6, being gone in SD-ura dextrose culture-medium later and adjusting initial concentration is OD₆₀₀= 0.1, the different pH gradients of setting 2,3,4,5,6,7,8 record the OD value of bacterium solution to 14 hours every 2h.

CsHT11 absorbs glucose as time growth is a linear process (Fig. 3) and CsHT11 is transported as the result is shown The optimal pH of glucose is 4-6 (Fig. 4).

5, CsHT11 gene subcellular localization

Using hexose transport PROTEIN C sHT11 gene, subcellular localization support C sHT11-pH7LIC5.0-ccdB is constructed Bacterium solution, is entered the lower epidermis of Ben's tobacco, laser by rc-N-eGFP, recombinant plasmid transformed Agrobacterium GV3101 by epidermal injection Observation discovery CsHT11 is located on cytoplasma membrane (Fig. 5) under Laser Scanning Confocal Microscope.

Sequence table

<110>Hua Zhong Agriculture University

<120>a kind of hexose transport albumen and its encoding gene and application

<160> 4

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1548

<212> DNA

<213>cucumber (Cucumis sativus L.)

<400> 1

atggctggag gaggatttgt tgctgaaggg agatcaggaa ggaactatga aggaggagtg 60

actggtttcg tgatagtcac atgtttggtg gctgccatgg gaggtcttct cttcggttat 120

gatcttggaa tctcaggtgg cgttacttca atgccaagtt tcttggatca gtttttccca 180

tctgttgtta agaagatgaa aggtgcccat gaaagtgagt actgtaaatt tgacagtgaa 240

ctccttactt tgtttacctc ttctctttac cttgctgcgc tcgtcgcttc cttcgccgcc 300

tctgtcatca cccgtaagtt tggtcggaaa ccatctatgt tctttggagg tctttccttc 360

ttgattggtt ccattcttaa tggtgttgct aatggcattg gtctcctcat cattggtcga 420

ttgttgcttg gtgttggagt tggttttgcc aatcagtctg tcccagttta cctctctgaa 480

atggcaccag ccaaaatcag aggagctctc aatatgggct tccaaatggc cattacaatc 540

ggcatcttgg tggcgagtct tgtcaacgtc ggtacatcca aaatcgaagg cggatgggga 600

tggagagtct cgctcgctct cgcctcagtc cccgccgtga tgatgaccat cggagccatc 660

ttccttccag atactcccaa ctccattctc gaaagaggat tcacagagaa ggccaagacg 720

atgctccaaa aagtccgcgg taccgataac gtggaagaag agttcaaaga ccttctggat 780

gcaagtgaag ctgcaaagaa agtagatcat ccatggacca acatactgaa gccacagtac 840

agacctcaac tcgtgatgtg caccataatc ccattcttcc aacaactcac cggcattaat 900

gttatcatgt tctacgctcc tgttctattc atgactctag gattcggcga cgacgcatct 960

ctaatctccg ccgtcatcag cggtggtgtc aacgtccttg caaccttagt ttccatcttc 1020

acagtggaca aattcggacg cagaatcctg ttcctcgaag gtggagtaca gatgttcatc 1080

tgccaaattc tagtaggaac tctaatcgga gttaagtttg gactgaacgg agaaggaacc 1140

ttatcgaaat tcgacgcaaa cctgattctg ttcctggtgt gcacatacgt agcggcattc 1200

gcttggtctt ggggaccatt gggttggttg gtaccaagtg aaatctgccc gctggaaatt 1260

cgatcagcag gacaagcaat aaacgtatca gtgaacatgt tcttcacgtt cataatcgca 1320

caagtgttcc tggcgatgct ttgccacatg aaattcggac tgttctactt cttcgcaggg 1380

tttgtgttga taatgacggt gtttatatat ttcttcttgc cggagacaaa gaatgtgccg 1440

attgaagaaa tgaatagagt gtggaaggcg cattggtttt gggggaagta cattcccgac 1500

gaggtgatca ttggcggccc tgtggggaag catgacgacg atgcttaa 1548

<210> 2

<211> 515

<212> RNA

<213>cucumber (Cucumis sativus L.)

<400> 2

<210> 3

<211> 28

<212> DNA

<213>artificial sequence (Artificial sequence)

<400> 3

atggctggag gaggatttgt tgctgaag 28

<210> 4

<211> 25

<212> DNA

<213>artificial sequence (Artificial sequence)

<400> 4

tcaagcatcg tcgtcatgct tcccc 25

Claims

1. hexose transport PROTEIN C sHT11 has the amino acid sequence as shown in sequence table SEQ ID NO:2.

2. encoding the gene of hexose transport PROTEIN C sHT11 described in claim 1.

3. gene according to claim 2, it is characterised in that: the gene has as shown in sequence table SEQ ID NO:1 Nucleotide sequence.

4. the recombinant expression carrier containing gene described in Claims 2 or 3.

5. the transgenic cell line containing gene described in Claims 2 or 3.

6. the recombinant bacterium containing gene described in Claims 2 or 3.

7. recombinant expression carrier according to claim 4, it is characterised in that: the recombinant expression carrier is pDR196's The recombinant expression carrier that gene described in Claims 2 or 3 obtains is inserted between multiple cloning sites.

8. recombinant bacterium according to claim 6, it is characterised in that: the recombinant bacterium is to carry described in Claims 2 or 3 The yeast of gene.

9. recombination table described in albumen described in claim 1 or gene described in claim 2 or 3 or claim 4 or 7 Microorganism is being improved to the application in hexose absorption up to carrier, and the hexose is glucose, fructose, galactolipin, mannose.

10. a kind of cultivate the method with the microorganism of high hexose absorbent properties, it is characterised in that include the following steps: right It is required that recombinant expression carrier described in 4 or 7 converts purpose microorganism, glucose, fructose, galactolipin, mannose absorbability are obtained The recombinant microorganism of the purpose microorganism can be higher than.