CN104073489B - A kind of DNA fragmentation inducing destination protein to express for uric acid and promoter related with application - Google Patents
A kind of DNA fragmentation inducing destination protein to express for uric acid and promoter related with application Download PDFInfo
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Abstract
The invention discloses a kind of DNA fragmentation inducing destination protein to express for uric acid and promoter related with application.The invention provides a kind of DNA molecular, its nucleotide sequence be in sequence table sequence 1 from 5 ' end 143-200 position nucleotide.DNA fragmentation containing above-mentioned DNA molecular is also the scope of protection of the invention.The experiment proves that, the present invention is by transforming E. coli lac promoter-35 district and-10th district, obtaining a new promoter, this promoter under the cooperation of uric acid modulin HucR and urate transporter YgfU, can utilize the expression of uric acid induction destination protein.Therefore the promoter of the present invention and correlated expression system can be used to carry out destination protein expression under the induction of uric acid, it is also possible to detect system in biotype uric acid.
Description
Technical field
The present invention relates to biological technical field, particularly relate to a kind of DNA fragmentation inducing destination protein to express for uric acid and promoter related with application.
Background technology
Expression of recombinant proteins system is the important research content of modern biotechnology, in field extensive uses such as food, pharmacy, detergent productions.Wherein, antibacterial heterologous expression system have bacterial growth for time short, thalli growth density is high, substrate is cheap, genetic background is clear, be appropriate to the significant advantages such as engineered strain transformation, especially escherichia expression system, it has also become one of expression system being most widely used.
Heterologous protein recombinant expressed, is usually required that after being placed in by desired protein coding sequences by technology such as molecular clonings and having stronger transcriptional capability promoter.At present, the studied report of promoter that existing multiple escherichia coli are suitable for.Wherein directly or indirectly by lac, tac, trc, T7 promoter of isopropyl-beta D-thio galactopyranoside regulation and control, by the tet promoter of Tetracycline regulation, by the Arabinose promoter (P of arabinose regulation and controlBAD), by the rhaP of rhamnose regulation and controlBADIt is employed Deng, the successful expression heterologous recombinant protein of number of different types.
Colibacillary lac operon is furtherd investigate.The transcriptional capability of Lac operon lac own is more weak, is not suitable for the great expression of recombiant protein.But the structure of many promoteres, is all transform to form in the function element that system handled by lactose.Such as, classical tac promoter and trc promoter are by-35th district of trp promoter and-10th district of lac promoter are combined and spliced forms, and have higher expression efficiency than lac promoter.T7 promoter then utilizes isopropyl-beta D-thio galactopyranoside to regulate and control the expression of t7 rna polymerase, and then opens T7 promoter systems expression downstream recombiant protein；This technology is developed series commercialization carrier, the successful expression heterologous protein of multiple separate sources by companies such as Novagen.
Sum up, a set of successful expression of recombinant proteins system, first require that the related elements of promoter can by Host Strains identification.And want successful expression heterologous recombinant protein, not requiring nothing more than promoter has higher transcriptional efficiency, also requires that the leakage expression of promoter is relatively low, and host cell more especially has the albumen of certain toxicity.Finally, considering from reducing production cost, the price of inducer own should be comparatively cheap, wide material sources, and is not degraded utilization by the metabolic system of Host Strains own.
The genome analysis having the extreme microorganism radioresistant cocci (Deinococcusradiodurans) of strong radiation hardness characteristic discloses the partial function assembly that this bacterium adverse-resistant characteristic is relevant.Wherein, the supposition uricase modulin (hypotheticaluricaseregulator of coding region dr1159 coding, HucR) research shows, HucR can form the DNA binding site sequence that albumen dimer is incorporated in the promoter region (hucO) of himself coded sequence and reverse uricase coded sequence；But after being combined with uric acid, HucR protein structure changes, and dissociates from DNA binding site；Therefore, HucR regulator control system becomes radioresistant cocci and changes according to environment uric acid situation, the system that the uricase that Effective Regulation HucR albumen itself and uric acid degraded are correlated with is expressed.Because uric acid is the compound that a kind of price is less expensive, and not utilizing for the metabolism of escherichia coli institute, this result of study imply that can be transformed this promoter, builds the escherichia coli expression of recombinant proteins system being inducer with uric acid.
Summary of the invention
It is an object of the present invention to provide a kind of DNA molecular.
DNA molecular provided by the invention, its nucleotide sequence be in sequence table sequence 1 from 5 ' end 143-200 position nucleotide.
Above-mentioned DNA molecular is also the scope of protection of the invention as the application in promoter.
DNA fragmentation containing above-mentioned DNA molecular is also the scope of protection of the invention.
Above-mentioned DNA fragmentation is made up of DNA fragmentation 1, destination protein gene and the DNA fragmentation 2 containing above-mentioned DNA molecular；
Described DNA fragmentation 1 be containing the promoter CP6 driving HucR encoding gene and YgfU encoding gene to express, HucR encoding gene, YgfU encoding gene fragment；
Above-mentioned DNA molecular is used for driving described destination protein gene expression.
The transcriptional orientation of above-mentioned HucR encoding gene and YgfU encoding gene is contrary with the transcriptional orientation of destination protein gene.
In above-mentioned DNA fragmentation, the coded sequence of the described promoter CP6 driving HucR encoding gene and YgfU encoding gene to express is the sequence 1 reverse complementary sequence from 5 ' end 3244-3303 nucleotide；
The coded sequence of described HucR encoding gene is the sequence 1 reverse complementary sequence from 5 ' end 2662-3224 nucleotide；
The coded sequence of described YgfU encoding gene is the sequence 1 reverse complementary sequence from 5 ' end 1197-2645 position nucleotide；
Described DNA fragmentation 1 is specially in sequence table sequence 1 from the shown double chain DNA molecule of 5 ' end 958-3303 position nucleotide；
Described DNA fragmentation 2 is specially in sequence table sequence 1 from the double chain DNA molecule shown in 5 ' end 143-279 position nucleotide.
In above-mentioned DNA fragmentation, described destination protein gene is red fluorescent protein gene, the coded sequence of described red fluorescent protein gene be in sequence table sequence 1 from 5 ' end 280-957 position nucleotide；
Described DNA fragmentation is that sequence 1 is from the double chain DNA molecule shown in 5 ' end 143-3303 nucleotide.
Also it is the scope of protection of the invention containing above-mentioned DNA molecular or containing the recombinant vector of above-mentioned DNA fragmentation, transgenic cell line or recombinant bacterium.
The nucleotides sequence of above-mentioned recombinant vector is classified as the sequence 1 in sequence table；
Above-mentioned recombinant bacterium is that described recombinant vector is proceeded to the recombinant bacterium obtained in Host Strains.In an embodiment of the present invention, above-mentioned Host Strains is specially escherichia coli DH10B.
Above-mentioned DNA molecular or above-mentioned DNA fragmentation or above-mentioned recombinant vector, transgenic cell line or recombinant bacterium induce the application in the expression of destination protein at uric acid.
The method that it is a further object to provide the expression of a kind of uric acid induction destination protein.
Method provided by the invention, is comprised the steps: to be imported in Host Strains the encoding gene of destination protein by above-mentioned recombinant vector, obtains recombinant bacterium；Ferment above-mentioned recombinant bacterium again in the culture medium containing uric acid, it is achieved uric acid induction destination protein.
Above-mentioned DNA molecular or above-mentioned DNA fragmentation or the application in utilizing destination protein expression monitoring uric acid concentration of above-mentioned recombinant vector, transgenic cell line or recombinant bacterium are also the scope of protection of the invention.
The experiment proves that, the present invention is by transforming E. coli lac promoter-35 district and-10th district, obtaining a new promoter, this promoter under the cooperation of uric acid modulin HucR and urate transporter YgfU, can utilize the expression of uric acid induction destination protein.Therefore the promoter of the present invention and correlated expression system can be used to carry out destination protein expression under the induction of uric acid.Present invention may also apply to uric acid biotype detection system.
Accompanying drawing explanation
Fig. 1 is the structural representation of SHY plasmid
Fig. 2 is uric acid inducible system induction curves
Detailed description of the invention
The experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, all commercially obtain.
The acquisition of embodiment 1, promoter A and the DNA fragmentation for uric acid induction
The DNA shown in sequence 1 in following embodiment is plasmid SHY, DNA shown in sequence 2 be nucleotide sequence sequence 1 and the control plasmid HY nucleotide sequence sequence 2 of control plasmid HY, SHY plasmid uniquely differ only in the lac promoter from 5 ' end 143-200 position nucleotide of sequence 2 is replaced with sequence 1 in sequence table from the promoter A shown in 5 ' end 143-200 position nucleotide.
The concrete construction method of above-mentioned plasmid is as follows:
1, the acquisition of uric acid modulin HucR and encoding gene thereof
Extract the genomic DNA of radioresistant cocci (Deinococcusradiodurans) (ATCC13939) and with it for template, carry out pcr amplification with 5 '-gcacatatgatgagcgcgcgcatggataac-3 ' (forward) and 5 '-agagagctcttaaacaccctgttcgaggc-3 ' (reverse) for primer.Pcr amplification condition is as follows: first 95 DEG C of denaturation 4min, then 95 DEG C of 45s, 55 DEG C of 30s, 72 DEG C of 30s, totally 30 circulations；Last 72 DEG C extend 10min.
Reclaim above-mentioned PCR product, carry out agarose gel electrophoresis detection, it is thus achieved that the band of 543bp (through order-checking, its coded sequence be in sequence table sequence 1 or 2 from the reverse complementary sequence of 5 ' end 2662-3224 nucleotide).
By PCR primer NdeI and the SacI double digestion of above-mentioned acquisition 543bp, DsRed carrier (Clontech company, production code member 632412) the carrier framework T of digestion products and the 3311bp through identical enzyme action4Ligase (purchased from precious biotech firm) 16 DEG C connects overnight, connects product and is transformed into escherichia coli MC1061 competent cell, it is thus achieved that recombinant bacterium.
Extract the plasmid of recombinant bacterium, this plasmid is for inserting DsRed carrier (existing constitutive promoter CP6 by sequence 1 or 2 in sequence table from the modulin HucR gene shown in the reverse complementary sequence of 5 ' end 2662-3224 nucleotide, after this promoter, insert modulin HucR gene) NdeI and SacI restriction enzyme site between the carrier that obtains, sequence verification is errorless, called after SH plasmid, it is the sequence 1 or 2 reverse complementary sequence from 5 ' end 3244-3303 nucleotide that this plasmid includes driving its coded sequence of promoter CP6(that HucR encoding gene is expressed), HucR encoding gene (its coded sequence is the sequence 1 or 2 reverse complementary sequence from 5 ' end 2662-3224 nucleotide).
In sequence table, sequence 1 is from the encoding gene that fragment shown in the reverse complementary sequence of 5 ' end 2662-3224 nucleotide is uric acid modulin HucR, and the aminoacid sequence of the uric acid modulin HucR of coding is sequence 3 in sequence table.
2, the acquisition of urate transporter YgfU and encoding gene and control plasmid HY
Extract escherichia coli MC1061(Casadaban, MJ&Cohen, SN (1980) AnalysisofgenecontrolsignalsbyDNAfusionandcloninginEsche richiacoli.J.Mol.Biol.138179-207；The public can obtain from Institute of Microorganism, Academia Sinica) genomic DNA and with it for template, carry out pcr amplification with 5 '-ttcgagctcaacaccctgttcgaggcccgc-3 ' (forward) and 5 '-gctgaagaaaaatgagcatggagaataagctagccca-3 ' (reverse) for primer.Pcr amplification condition is as follows: first 95 DEG C of denaturation 4min, then 95 DEG C of 45s, 55 DEG C of 30s, 72 DEG C of 1min30s, totally 30 circulations；Last 72 DEG C extend 10min.
Reclaim above-mentioned PCR product, carry out agarose gel electrophoresis detection, it is thus achieved that the band (for sequence in sequence table 1 from the transport protein YgfU encoding gene shown in the reverse complementary sequence of 5 ' end 1197-2645 position nucleotide) of 1449bp.
By PCR primer NheI and the SacI double digestion of the 1449bp size of above-mentioned acquisition, the SH carrier framework T of digestion products and the 3941bp through identical enzyme action4Ligase (purchased from precious biotech firm) 16 DEG C connects overnight, connects product and is transformed into escherichia coli MC1061 competent cell, it is thus achieved that recombinant bacterium.
Extract the plasmid of recombinant bacterium, this plasmid is for inserting the SacI enzyme of SH carrier by sequence in sequence table 1 from the transport protein YgfU encoding gene shown in the reverse complementary sequence of 5 ' end 1197-2645 position nucleotide and NheI cuts the carrier obtained between site, sequence verification is errorless, (ygfU gene closelys follow hucR gene to called after HY plasmid, also utilizes CP6 promoter to express.
nullHY plasmid includes comparison DNA fragmentation，It is the sequence 1 or 2 reverse complementary sequence from 5 ' end 3244-3303 nucleotide that comparison DNA fragmentation includes driving its coded sequence of promoter CP6(that HucR encoding gene and YgfU encoding gene are expressed)、HucR encoding gene (its coded sequence is the sequence 1 or 2 reverse complementary sequence from 5 ' end 2662-3224 nucleotide)、YgfU encoding gene (its coded sequence is the sequence 1 or 2 reverse complementary sequence from 5 ' end 1197-2645 position nucleotide)、Red fluorescent protein RFP gene (its coded sequence be in sequence table sequence 1 or 2 from 5 ' end 280-957)、Its coded sequence of Lac operon lac(driving red fluorescent protein RFP gene expression is that sequence 2 is from 5 ' end 143-200 position nucleotide).
Comparison DNA fragmentation is formed by containing the promoter CP6, HucR encoding gene, the DNA fragmentation 1 of YgfU encoding gene, red fluorescent protein RFP gene and the DNA fragmentation 2 containing the Lac operon lac driving red fluorescent protein RFP gene expression that drive HucR encoding gene and YgfU encoding gene to express；Wherein, DNA fragmentation 1 be in sequence table sequence 2 from the shown double chain DNA molecule of 5 ' end 958-3303 position nucleotide, red fluorescent protein RFP gene coded sequence be in sequence table sequence 1 or 2 from 5 ' end 280-957, DNA fragmentation 2 for sequence in sequence table 2 from the double chain DNA molecule shown in 5 ' end 143-279 position nucleotide.
The nucleotides sequence of HY plasmid is classified as the sequence 2 in sequence table, wherein comparison DNA fragmentation be in sequence table sequence 2 from the double chain DNA molecule shown in 5 ' end 143-3303 nucleotide.
In sequence table, sequence 1 or 2 is from the encoding gene that fragment is urate transporter YgfU shown in the reverse complementary sequence of 5 ' end 1197-2645 position nucleotide, and the aminoacid sequence of the urate transporter YgfU of coding is sequence 4 in sequence table.
3, promoter A that genes of interest (red fluorescent protein gene) expresses is driven, for the DNA fragmentation of uric acid induction and the acquisition of recombinant vector SHY
In radioresistant cocci genome, uric acid modulin HucR DNA binding sequence row in hucO are confirmed in experiment.In order to transform-35th district and-10th district of the promoter lac in HY plasmid, add new fragment when designing following primer, obtain new promoter.
With 5 '-atctaagtatatgttGtgtggaaccgatttaataaaacaa-3 ' (forward) and 5 '-gtctacctatgtaaaGcctggggtgcctaatg-3 ' (reverse) is primer, the above-mentioned 2 HY plasmids obtained are that template carries out pcr amplification, by HY plasmid linearization.Pcr amplification condition is as follows: first 95 DEG C of denaturation 4min, then 95 DEG C of 45s, 55 DEG C of 30s, 72 DEG C of 6min, totally 30 circulations；Last 72 DEG C extend 10min..
Reclaim above-mentioned PCR product, carry out agarose gel electrophoresis detection, it is thus achieved that the band of 5404bp.
By above-mentioned 5404bp linearization plasmid fragment T4Ligase (purchased from precious biotech firm) 16 DEG C connects overnight, connects product and is transformed into escherichia coli MC1061 competent cell, it is thus achieved that recombinant bacterium.
Extracting the plasmid of recombinant bacterium, the nucleotides sequence of this plasmid is classified as the sequence 1 in sequence table, and called after SHY plasmid, for recombinant vector (its structural representation is as it is shown in figure 1, the promoter wherein regulated and controled by uric acid is drive the promoter A of destination protein gene expression).
nullIn SHY plasmid containing for uric acid induction DNA fragmentation，Including driving its coded sequence of promoter CP6(that HucR encoding gene and YgfU encoding gene are expressed for the DNA fragmentation of uric acid induction is the sequence 1 or 2 reverse complementary sequence from 5 ' end 3244-3303 nucleotide)、HucR encoding gene (its coded sequence is the sequence 1 or 2 reverse complementary sequence from 5 ' end 2662-3224 nucleotide)、YgfU encoding gene (its coded sequence is the sequence 1 or 2 reverse complementary sequence from 5 ' end 1197-2645 position nucleotide)、Red fluorescence RFP protein gene (its coded sequence be in sequence table sequence 1 or 2 from 5 ' end 280-957)、Its coded sequence of promoter A(driving red fluorescent protein RFP gene expression is that sequence 1 is from 5 ' end 143-200 position nucleotide).
DNA fragmentation for uric acid induction is formed by containing the promoter CP6, HucR encoding gene, the DNA fragmentation 1 of YgfU encoding gene, red fluorescent protein RFP gene and the DNA fragmentation 2 containing the promoter A driving red fluorescent protein RFP gene expression that drive HucR encoding gene and YgfU encoding gene to express；Wherein, DNA fragmentation 1 be in sequence table sequence 1 from the shown double chain DNA molecule of 5 ' end 958-3303 position nucleotide, red fluorescent protein RFP gene coded sequence be in sequence table sequence 1 or 2 from 5 ' end 280-957, DNA fragmentation 2 for sequence in sequence table 1 from the double chain DNA molecule shown in 5 ' end 143-279 position nucleotide.
It is classified as sequence 1 in sequence table from the double chain DNA molecule (namely including after transformation by uric acid regulation and control promoter, reporter gene, modulin HucR and transport protein YgfU by constitutive promoter regulating and expressing) shown in 5 ' end 143-3303 nucleotide for the nucleotides sequence of DNA fragmentation of uric acid induction.
The nucleotide sequence sequence 1 of SHY plasmid and control plasmid HY nucleotide sequence sequence 2 uniquely differ only in by sequence 2 from the promoter lac shown in 5 ' end 143-200 position nucleotide replace with sequence 1 in sequence table from the promoter A shown in 5 ' end 143-200 position nucleotide.
The application in uric acid induction destination gene expression of embodiment 2, promoter A and the DNA fragmentation for uric acid induction
The recombinant vector SHY obtained by embodiment 1 and control plasmid HY is proceeded in escherichia coli DH10B respectively, obtains the recombinant bacterium DH10B/SHY containing SHY and to extract plasmid sequence verification respectively containing the recombinant bacterium DH10B/HY(of HY correct).
Single bacterium colony of picking recombinant bacterium DH10B/SHY and recombinant bacterium DH10B/HY, at LB culture medium (the tryptone 10g/L adding ammonia benzyl antibiotic resistance, NaCl10g/L, yeast extract 5g/L, adding antibiotic ammonia benzyl mycin to 100 μ g/mL) 37 DEG C of shaking table overnight incubation (12h) to bacteria concentrations reach capacity (OD600 is 3.5), obtain seed liquor；
Seed liquor accesses induction 12h in the uric acid culture medium of variable concentrations according to inoculum concentration 1%, and condition of culture is 37 DEG C of shaking tables (250rpm).Terminating to induction time, take out culture, centrifugal thalline of collecting abandons supernatant (3000g, 10min)；Again with phosphate buffer (100mM, pH6.0) resuspended thalline, draw 200 μ L thalline and test bacteria concentration OD with microplate reader600And red fluorescent protein reading.
nullThe uric acid culture medium of above-mentioned variable concentrations is prepared as follows: by tryptone、NaCl、Yeast extract、Ammonia benzyl mycin、Uric acid and pH7.0、Concentration is 3-morpholine propane sulfonic acid (MOPS) buffer mother solution and the water mixing of 500mM，The wherein final concentration of 10g/L of tryptone、The final concentration of 10g/L of NaCl、The final concentration of 5g/L of yeast extract、The final concentration of 100 μ g/mL of ammonia benzyl mycin、The final concentration of 17mM of 3-morpholine propane sulfonic acid (MOPS) buffer，Uric acid final concentration respectively 0mM in the uric acid culture medium of variable concentrations、0.002mM、0.005mM、0.006mM、0.008mM、0.01mM、0.015mM、0.02mM、0.03mM、0.04mM and 0.05mM.
The induction result of recombinant bacterium DH10B/SHY is as in figure 2 it is shown, vertical coordinate is reporter gene red fluorescent protein fold induction (ratio for the dialuric acid induced reporter gene red fluorescent protein reading of uric acid induced reporter gene red fluorescent protein reading/not)；Can be seen that, in culture medium in the final concentration of 0mM of uric acid, 0.002mM, 0.005mM, 0.006mM, 0.008mM, 0.01mM, 0.015mM, 0.02mM, 0.03mM, 0.04mM and 0.05mM, after recombinant bacterium DH10B/SHY growth induction expression protein, reporter gene red fluorescent protein reading respectively 168,1539,7506,15726,20247,27946,40416,52698,62883,70139,72818；The ratio respectively 1,9.16,44.67,93.61,120.51,166.34,240.57,313.67,374.30,417.49,433.44 times of the dialuric acid induced reporter gene red fluorescent protein reading of uric acid induced reporter gene red fluorescent protein reading/not.Test result shows, along with uric acid concentration increases, reporter gene red fluorescent protein reading increases.Illustrate promoter A, containing promoter A for uric acid induction DNA fragmentation, SHY carrier destination protein red fluorescent protein can be made under the induction of uric acid to express.
The induction result of recombinant bacterium DH10B/HY is as follows: in the culture medium in the final concentration of 0mM of uric acid, 0.002mM, 0.005mM, 0.006mM, 0.008mM, 0.01mM, 0.015mM, 0.02mM, 0.03mM, 0.04mM and 0.05mM, after recombinant bacterium DH10B/HY growth induction expression protein, reporter gene red fluorescent protein reading is 80-160 substantially；Dialuric acid induction (0mM) reporter gene red fluorescent protein reading is not 153；It can therefore be seen that uric acid induced reporter gene red fluorescent protein reading/ratio of not dialuric acid induced reporter gene red fluorescent protein reading substantially within 1, namely do not induced by uric acid.
The induction result of recombinant bacterium DH10B/SHY shows, when uric acid concentration is lower than 0.01mM, uric acid induced reporter gene red fluorescent protein fold induction is linear with uric acid concentration, formula is that (wherein y is red fluorescent protein fold induction to y=11946x-11.77, x is uric acid concentration, this standard curve standard error square is up to 0.97, and linearisation is good).It is therefore possible to use the carrier of the present invention, promoter or fragment are for extraneous uric acid concentration monitoring.
The above results can be seen that, compared with control plasmid HY, SHY plasmid can express destination protein better under uric acid is induced, and SHY plasmid and control plasmid HY uniquely differ only in the difference of promoter, illustrate that the promoter A of SHY plasmid has the function expressing destination protein better under uric acid is induced.
Claims (9)
1. a DNA molecular, its nucleotide sequence be in sequence table sequence 1 from 5 ' end 143-200 position nucleotide.
2. DNA molecular described in claim 1 is as the application in promoter.
3. contain the DNA fragmentation of DNA molecular described in claim 1；Described DNA fragmentation is made up of DNA fragmentation 1, destination protein gene and the DNA fragmentation 2 containing DNA molecular described in claim 1；
Described DNA fragmentation 1 be containing the promoter CP6 driving HucR encoding gene and YgfU encoding gene to express, HucR encoding gene, YgfU encoding gene fragment；
DNA molecular described in claim 1 is used for driving described destination protein gene expression.
4. DNA fragmentation according to claim 3, it is characterised in that:
The coded sequence of the described promoter CP6 driving HucR encoding gene and YgfU encoding gene to express is the sequence 1 reverse complementary sequence from 5 ' end 3244-3303 nucleotide；
The coded sequence of described HucR encoding gene is the sequence 1 reverse complementary sequence from 5 ' end 2662-3224 nucleotide；
The coded sequence of described YgfU encoding gene is the sequence 1 reverse complementary sequence from 5 ' end 1197-2645 position nucleotide；
Described DNA fragmentation 1 is specially in sequence table sequence 1 from the shown double chain DNA molecule of 5 ' end 958-3303 position nucleotide；
Described DNA fragmentation 2 is specially in sequence table sequence 1 from the double chain DNA molecule shown in 5 ' end 143-279 position nucleotide.
5. the DNA fragmentation according to claim 3 or 4, it is characterised in that: described destination protein gene is red fluorescent protein gene, the coded sequence of described red fluorescent protein gene be in sequence table sequence 1 from 5 ' end 280-957 position nucleotide；
Described DNA fragmentation is that sequence 1 is from the double chain DNA molecule shown in 5 ' end 143-3303 nucleotide.
6. contain the DNA molecular described in claim 1 or containing the recombinant vector of arbitrary described DNA fragmentation, transgenic cell line or recombinant bacterium in claim 3-5；
Described recombinant bacterium is that described recombinant vector is proceeded to the recombinant bacterium obtained in Host Strains.
7. DNA molecular described in claim 1 or in claim 3-5 recombinant vector described in arbitrary described DNA fragmentation or claim 6, transgenic cell line or recombinant bacterium induce the application in the expression of destination protein at uric acid.
8. the method for the expression of a uric acid induction destination protein, it is characterised in that: the encoding gene of destination protein is imported in Host Strains by the recombinant vector described in claim 6, obtains recombinant bacterium；Ferment described recombinant bacterium again in the culture medium containing uric acid, it is achieved uric acid induction destination protein.
9. DNA molecular described in claim 1 or recombinant vector described in arbitrary described DNA fragmentation or claim 6, transgenic cell line or recombinant bacterium application in utilizing destination protein expression monitoring uric acid concentration in claim 3-5.
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