CN103013941A - Preparation method and application of sarcosine oxidase - Google Patents
Preparation method and application of sarcosine oxidase Download PDFInfo
- Publication number
- CN103013941A CN103013941A CN2012105784815A CN201210578481A CN103013941A CN 103013941 A CN103013941 A CN 103013941A CN 2012105784815 A CN2012105784815 A CN 2012105784815A CN 201210578481 A CN201210578481 A CN 201210578481A CN 103013941 A CN103013941 A CN 103013941A
- Authority
- CN
- China
- Prior art keywords
- sarcosine oxidase
- sarcosine
- prepare
- expression
- expression vector
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention discloses a preparation method of sarcosine oxidase. The method comprises the following steps: transforming a whole-gene synthesis humanization sarcosine oxidase coding deoxyribonucleic acid sequence, and removing escherichia coli rare codon; inserting the transformed sarcosine oxidase coding deoxyribonucleic acid sequence into an expression carrier, and carrying out transformation and expression by using an escherichia coli strain; inoculating the strain subjected to transformation and expression into a luria-bertani (LB) culture medium, and carrying out isopropyl-beta-d-thiogalactoside (IPTG) induction; and breaking the obtained thallus, centrifugally separating and purifying through an Ni-nitrilotriacetic acid (NTA) column. The method is simple in sarcosine oxidase preparation operation, low in preparation cost, high in finished product yield, high in purity and high in biological activity, and can be used as a diagnostic reagent for sarcosine assay kit research.
Description
Technical field
The invention belongs to genetically engineered and protein expression field, relate to and a kind ofly efficiently prepare the method for sarcosine oxidase with biotechnology, product can be used for producing the sarkosine detection kit.
Background technology
Univ Michigan-Ann Arbor USA Ann Arbor branch school Xin Laiyan (Chinnaiyan) etc. is published in " nature " magazine on February 12nd, 2009, and (Nature 2009, research 457:910) claims, a kind of sarkosine (sarcosine by name, also the name sarcosine, the metabolite of glycine) material can effectively reflect the aggressive of prostate cancer, distinguishes the growth behavior (slowly growth or highly invasion and attack) of cancer cells.In addition two important discoveries of this research are: sarkosine has participated in prostatic Carcinogenesis, is one of arch-criminal of cell carcinogenesis; The accuracy that the urine sarkosine detects diagnosing prostate cancer is better than present blood prostate specific antigen (PSA) detection of commonly using clinically.
The investigator finds to have at least the level of 6 kinds of metabolites to have significant difference in good malignant prostate tissue, and is wherein remarkable with sarkosine again.The sarkosine level is extremely low in benign prostate, presents significantly to raise in the limitation prostate cancer, then rises De Genggao in metastatic prostate cancer.
At present mainly contain liquid chromatography or chromatography of gases and mass spectrometry detection method, sarcosine oxidase fluorescence detection, capillary electrophoresis electrochemical light-emitting detection method, sarcosine oxidase Spectrophotometric Assays method for detection of the method for sarkosine.
Although chromatograph-mass spectrometer coupling detection method detection sensitivity, accuracy, precision are all higher, because the chromatography-mass spectroscopy apparatus expensive, testing cost is higher, and detection speed is slow, is difficult to mass detection, and the complicated operation common people are difficult to grasp.Therefore, often be applied to scientific research, be difficult to applying clinical large sample amount diagnostic detection.
Sarcosine oxidase fluorescence detection, its detection sensitivity of capillary electrophoresis electrochemical light-emitting detection method, accuracy, precision is a little less than the chromatograph-mass spectrometer coupling detection method, but also can satisfy clinical demand.The testing cost of these two kinds of methods reduces greatly than the chromatograph-mass spectrometer coupling detection method, but still need purchase special proofing unit when detecting, such as Special Equipments such as fluorimetric detector, capillary photoelectrophoresis instrument.And manually-operated part is very high, detects and still has certain difficulty being applied to routine clinical.
Sarcosine oxidase Spectrophotometric Assays method, its testing cost is low, can be applicable to present any automatic clinical chemistry analyzer, can be applicable to the detection needs of clinical high speed large sample amount.
Detect principle:
Having the quinone imides coloring matter to generate after reaction finishes causes the variation of absorbancy to be directly proportional with the concentration of sarkosine.
Summary of the invention
For the problem that prior art exists, the object of the present invention is to provide a kind of simple to operate, preparation cost is cheap, the method for preparing sarcosine oxidase that success ratio is high.
1. for achieving the above object, a kind of method for preparing sarcosine oxidase of the present invention, the method comprises:
1) transforms the synthetic humanization sarcosine oxidase coding gene sequence of also full gene, remove the intestinal bacteria rare codon;
2) with step 1) in improved sarcosine oxidase coding gene sequence insert in the expression vector, and transform with coli strain and to express;
3) will transform the expression inoculation in the LB substratum, IPTG induces;
4) the broken bacterium of gained thalline, the Ni-NTA column purification is crossed in centrifugation;
Wherein, the synthetic humanization sarcosine oxidase coding gene sequence of improved full gene is step 1):
ATGGCTTCTT?CTACTGGTGA?TCGTTCTCAG?GCTGTTCGTC?ATGGTCTGCG?TGCGAAAGTT
CTGACCCTGG?ACGGCATGAA?CCCGCGTGTT?CGTCGCGTCG?AATACGCCGT?GCGTGGCCCG
ATCGTTCAGC?GTGCTCTGGA?ACTGGAACAA?GAACTGCGTC?AGGGCGTTAA?AAAGCCGTTC
ACCGAGGTTA?TCCGCGCCAA?CATCGGCGAT?GCCCAGGCTA?TGGGTCAGCG?TCCTATCACC
TTCCTGCGCC?AGGTCCTGGC?GCTGTGCGTA?AACCCAGACC?TGCTGAGCTC?TCCGAACTTC
CCGGACGACG?CTAAGAAACG?TGCGGAACGC?ATCCTGCAGG?CATGCGGTGG?CCACAGCCTG
GGCGCTTACA?GCGTTTCCTC?CGGTATCCAG?CTGATTCGTG?AAGATGTTGC?CCGTTACATC
GAGCGCCGTG?ATGGCGGTAT?CCCGGCGGAC?CCTAACAACG?TATTCCTGTC?TACCGGTGCG
TCCGATGCCA?TTGTTACCGT?TCTGAAGCTG?CTGGTGGCTG?GCGAGGGTCA?CACCCGTACC
GGTGTCCTGA?TTCCGATTCC?ACAGTATCCG?CTGTATTCTG?CTACTCTGGC?TGAACTGGGT
GCGGTGCAGG?TAGACTATTA?CCTGGATGAG?GAGCGCGCTT?GGGCGCTGGA?CGTCGCAGAG
CTGCACCGCG?CACTGGGCCA?GGCACGTGAC?CACTGCCGTC?CGCGTGCGCT?GTGCGTGATT
AACCCGGGCA?ACCCGACGGG?CCAAGTTCAG?ACCCGTGAAT?GCATCGAAGC?TGTGATTCGT
TTTGCTTTCG?AAGAACGTCT?GTTCCTGCTG?GCGGACGAAG?TTTATCAGGA?CAACGTTTAC
GCAGCCGGTA?GCCAGTTTCA?TTCTTTCAAG?AAAGTTCTGA?TGGAAATGGG?TCCGCCGTAT
GCGGGTCAGC?AAGAACTGGC?ATCCTTCCAC?TCTACCTCCA?AAGGTTATAT?GGGTGAGTGT
GGCTTCCGTG?GTGGTTATGT?TGAAGTAGTT?AATATGGACG?CGGCTGTGCA?GCAGCAGATG
CTGAAGCTGA?TGTCTGTGCG?CCTGTGCCCG?CCTGTGCCGG?GTCAAGCCCT?GCTGGATCTG
GTTGTGTCTC?CGCCTGCTCC?GACTGATCCG?TCCTTCGCAC?AATTCCAGGC?TGAGAAACAG
GCCGTTCTGG?CGGAACTGGC?AGCGAAAGCC?AAACTGACGG?AACAAGTGTT?CAACGAAGCG
CCGGGTATCT?CCTGCAATCC?GGTACAAGGC?GCGATGTATA?GCTTCCCGCG?CGTTCAGCTG
CCGCCACGCG?CAGTAGAACG?TGCACAGGAA?CTGGGCCTGG?CTCCGGACAT?GTTCTTCTGT
CTGCGCCTGC?TGGAAGAAAC?CGGTATTTGC?GTTGTACCTG?GCAGCGGCTT?CGGTCAACGT
GAAGGTACTT?ACCACTTCCG?TATGACCATC?CTGCCTCCGC?TGGAAAAACT?GCGTCTGCTG
CTGGAAAAGC?TGTCCCGTTT?TCACGCGAAA?TTCACTCTGG?AATATTCTTA?G
Further, expression vector is step 2): pRSF Duet, pET express series.
Further, coli strain is step 2): BL21 (DE3), Rosetta (DE3).
Further, described BL21 (DE3) bacterial strain is used for efficiently expressing the gene that is cloned in the expression vector that contains the phage t7 promotor.
Further, lambda particles phage DE3 contains the T7 phage rna polymerase in the district, and this district is integrated in and forms described BL21 (DE3) on the karyomit(e) of BL21.
Further, broken bacterium method is broken, the ultrasonic or N,O-Diacetylmuramidase enzymolysis of high pressure step 4).
A kind of application of method in diagnostic reagent for preparing as claimed in claim 1 sarcosine oxidase.
Beneficial effect of the present invention is: the present invention prepares that sarcosine oxidase operation is very simple, and preparation cost is low, and finished product yield is high, and purity is high and have high biological activity, can be used as the development that diagnostic reagent is used for the sarkosine detection kit.
Description of drawings
The SAO pRSF that Fig. 1 illustrates structure cuts with NdeI and Xho I enzyme, obtains the Insert Fragment of about 1.2kb, shows insertion vector of goal gene, successfully constructs.
Fig. 2 illustrates the SDS-PAGE electrophorogram of the SAO finished product of purifying, and product purity can reach more than 95%.
Embodiment
Below, with reference to the accompanying drawings, the present invention is more fully illustrated, shown in the drawings of exemplary embodiment of the present invention.Yet the present invention can be presented as multiple multi-form, and should not be construed as the exemplary embodiment that is confined to narrate here.But, these embodiment are provided, thereby make the present invention comprehensively with complete, and scope of the present invention is fully conveyed to those of ordinary skill in the art.
The present invention has set up a kind of method for preparing sarcosine oxidase simple to operate, rapid, lower-cost.The principle of this method mainly is: the sarcosine oxidase encoding gene after will optimizing inserts the expression vector that builds, and induces it to efficiently express in intestinal bacteria, and the method is specially:
1. the structure of sarcosine oxidase expression vector:
Utilize NdeI and Xho I site to be cloned in the PRSF Duet carrier by genetic engineering means, and conversion intestinal bacteria TOP10 bacterial strain, several transformed clones of picking extract plasmid, identify whether insertion vector (shown in Figure 1) of goal gene by digestion with restriction enzyme, choose the plasmid that inserts goal gene and carry out determined dna sequence, the result shows that the sarcosine oxidase gene successfully is cloned in the pRSF Duet carrier, and we are with its called after SAO pRSF.
The synthetic humanization sarcosine oxidase coding gene sequence of improved full gene is:
ATGGCTTCTT?CTACTGGTGA?TCGTTCTCAG?GCTGTTCGTC?ATGGTCTGCG?TGCGAAAGTT
CTGACCCTGG?ACGGCATGAA?CCCGCGTGTT?CGTCGCGTCG?AATACGCCGT?GCGTGGCCCG
ATCGTTCAGC?GTGCTCTGGA?ACTGGAACAA?GAACTGCGTC?AGGGCGTTAA?AAAGCCGTTC
ACCGAGGTTA?TCCGCGCCAA?CATCGGCGAT?GCCCAGGCTA?TGGGTCAGCG?TCCTATCACC
TTCCTGCGCC?AGGTCCTGGC?GCTGTGCGTA?AACCCAGACC?TGCTGAGCTC?TCCGAACTTC
CCGGACGACG?CTAAGAAACG?TGCGGAACGC?ATCCTGCAGG?CATGCGGTGG?CCACAGCCTG
GGCGCTTACA?GCGTTTCCTC?CGGTATCCAG?CTGATTCGTG?AAGATGTTGC?CCGTTACATC
GAGCGCCGTG?ATGGCGGTAT?CCCGGCGGAC?CCTAACAACG?TATTCCTGTC?TACCGGTGCG
TCCGATGCCA?TTGTTACCGT?TCTGAAGCTG?CTGGTGGCTG?GCGAGGGTCA?CACCCGTACC
GGTGTCCTGA?TTCCGATTCC?ACAGTATCCG?CTGTATTCTG?CTACTCTGGC?TGAACTGGGT
GCGGTGCAGG?TAGACTATTA?CCTGGATGAG?GAGCGCGCTT?GGGCGCTGGA?CGTCGCAGAG
CTGCACCGCG?CACTGGGCCA?GGCACGTGAC?CACTGCCGTC?CGCGTGCGCT?GTGCGTGATT
AACCCGGGCA?ACCCGACGGG?CCAAGTTCAG?ACCCGTGAAT?GCATCGAAGC?TGTGATTCGT
TTTGCTTTCG?AAGAACGTCT?GTTCCTGCTG?GCGGACGAAG?TTTATCAGGA?CAACGTTTAC
GCAGCCGGTA?GCCAGTTTCA?TTCTTTCAAG?AAAGTTCTGA?TGGAAATGGG?TCCGCCGTAT
GCGGGTCAGC?AAGAACTGGC?ATCCTTCCAC?TCTACCTCCA?AAGGTTATAT?GGGTGAGTGT
GGCTTCCGTG?GTGGTTATGT?TGAAGTAGTT?AATATGGACG?CGGCTGTGCA?GCAGCAGATG
CTGAAGCTGA?TGTCTGTGCG?CCTGTGCCCG?CCTGTGCCGG?GTCAAGCCCT?GCTGGATCTG
GTTGTGTCTC?CGCCTGCTCC?GACTGATCCG?TCCTTCGCAC?AATTCCAGGC?TGAGAAACAG
GCCGTTCTGG?CGGAACTGGC?AGCGAAAGCC?AAACTGACGG?AACAAGTGTT?CAACGAAGCG
CCGGGTATCT?CCTGCAATCC?GGTACAAGGC?GCGATGTATA?GCTTCCCGCG?CGTTCAGCTG
CCGCCACGCG?CAGTAGAACG?TGCACAGGAA?CTGGGCCTGG?CTCCGGACAT?GTTCTTCTGT
CTGCGCCTGC?TGGAAGAAAC?CGGTATTTGC?GTTGTACCTG?GCAGCGGCTT?CGGTCAACGT
GAAGGTACTT?ACCACTTCCG?TATGACCATC?CTGCCTCCGC?TGGAAAAACT?GCGTCTGCTG
CTGGAAAAGC?TGTCCCGTTT?TCACGCGAAA?TTCACTCTGG?AATATTCTTA?G
2. the expression of sarcosine oxidase:
1) take BL21 (DE3) as Host Strains, SAO pRSF is transformed, picking list bacterium colony spends the night in the 3mlLB culture medium culturing;
2) with 1: 500 ratio bacterium liquid is forwarded to 400ml LB substratum, constant temperature is at 37 ℃, and 230rpm is cultured to OD600=0.4-0.6;
3) add IPTG and induce, it is 0.5mM that IPTG induces final concentration, keeps expressing after 8 hours under 25 ℃ of conditions receiving bacterium.
BL21 (DE3) bacterial strain is used for efficiently expressing the gene that is cloned in the expression vector (such as pET series) that contains the phage t7 promotor.Bacillus coli communis does not have the t7 RNA polymerase, so can not express the gene on those carriers.Lambda particles phage DE3 contains the T7 phage rna polymerase in the district, and this district is integrated on the karyomit(e) of BL21, just is BL21 (DE3).
3. the purifying of sarcosine oxidase:
1) get 400ml nutrient solution 3000rpm centrifugation and collect thalline after 10 minutes, thalline is resuspended in (50mM Na-PO in the 10ml protein extraction damping fluid
4, pH8.0,500mM NaCl, 10mMImidazle), the 15MPa height crushes bacterium, 12,000rpm, 4 ℃ of centrifugal 10min, supernatant liquor and Ni-NTA were hatched 1 hour on ice;
2) cross the Ni-NTA affinity column, with protein extraction buffer washing, use at last 250mM Imidazle wash-out, collect elutriant.
3) the albumen elutriant is crossed the G50 post and is removed Imidazle, can obtain like this SAO (shown in Figure 2) of 95% above purity.
4. sarcosine oxidase determination of activity:
The sterling of preparation sarcosine oxidase is measured active with Olympus AU5400 biochemical instruments, enzyme work can reach 30000U/L, and frozen 6 months of-20 ℃ of glycerine, and enzyme is lived and substantially remained unchanged.
The above-mentioned method for preparing sarcosine oxidase can be used as diagnostic reagent, is used for the development of sarkosine detection kit.
Unless specifically defined, it is known term in the relevant technologies field that the present invention describes used term.The chemical symbol of standard and dummy suffix notation can with its full name Alternate.
Unless special indicating, the present invention uses but clearly sets forth or simple technology and the method for setting forth refers to the normally used technology of the art and method, can carry out according to technology well known in the art and method.The use of test kit is to carry out according to the specification sheets that manufacturers or supplier provide.
Claims (7)
1. method for preparing sarcosine oxidase is characterized in that the method comprises:
1) transforms the synthetic humanization sarcosine oxidase coding gene sequence of also full gene, remove the intestinal bacteria rare codon;
2) with step 1) in improved sarcosine oxidase coding gene sequence insert in the expression vector, and transform with coli strain and to express;
3) will transform the expression inoculation in the LB substratum, IPTG induces;
4) the broken bacterium of gained thalline, the Ni-NTA column purification is crossed in centrifugation;
Wherein, the synthetic humanization sarcosine oxidase coding gene sequence of improved full gene is step 1):
ATGGCTTCTT?CTACTGGTGA?TCGTTCTCAG?GCTGTTCGTC?ATGGTCTGCG?TGCGAAAGTT
CTGACCCTGG?ACGGCATGAA?CCCGCGTGTT?CGTCGCGTCG?AATACGCCGT?GCGTGGCCCG
ATCGTTCAGC?GTGCTCTGGA?ACTGGAACAA?GAACTGCGTC?AGGGCGTTAA?AAAGCCGTTC
ACCGAGGTTA?TCCGCGCCAA?CATCGGCGAT?GCCCAGGCTA?TGGGTCAGCG?TCCTATCACC
TTCCTGCGCC?AGGTCCTGGC?GCTGTGCGTA?AACCCAGACC?TGCTGAGCTC?TCCGAACTTC
CCGGACGACG?CTAAGAAACG?TGCGGAACGC?ATCCTGCAGG?CATGCGGTGG?CCACAGCCTG
GGCGCTTACA?GCGTTTCCTC?CGGTATCCAG?CTGATTCGTG?AAGATGTTGC?CCGTTACATC
GAGCGCCGTG?ATGGCGGTAT?CCCGGCGGAC?CCTAACAACG?TATTCCTGTC?TACCGGTGCG
TCCGATGCCA?TTGTTACCGT?TCTGAAGCTG?CTGGTGGCTG?GCGAGGGTCA?CACCCGTACC
GGTGTCCTGA?TTCCGATTCC?ACAGTATCCG?CTGTATTCTG?CTACTCTGGC?TGAACTGGGT
GCGGTGCAGG?TAGACTATTA?CCTGGATGAG?GAGCGCGCTT?GGGCGCTGGA?CGTCGCAGAG
CTGCACCGCG?CACTGGGCCA?GGCACGTGAC?CACTGCCGTC?CGCGTGCGCT?GTGCGTGATT
AACCCGGGCA?ACCCGACGGG?CCAAGTTCAG?ACCCGTGAAT?GCATCGAAGC?TGTGATTCGT
TTTGCTTTCG?AAGAACGTCT?GTTCCTGCTG?GCGGACGAAG?TTTATCAGGA?CAACGTTTAC
GCAGCCGGTA?GCCAGTTTCA?TTCTTTCAAG?AAAGTTCTGA?TGGAAATGGG?TCCGCCGTAT
GCGGGTCAGC?AAGAACTGGC?ATCCTTCCAC?TCTACCTCCA?AAGGTTATAT?GGGTGAGTGT
GGCTTCCGTG?GTGGTTATGT?TGAAGTAGTT?AATATGGACG?CGGCTGTGCA?GCAGCAGATG
CTGAAGCTGA?TGTCTGTGCG?CCTGTGCCCG?CCTGTGCCGG?GTCAAGCCCT?GCTGGATCTG
GTTGTGTCTC?CGCCTGCTCC?GACTGATCCG?TCCTTCGCAC?AATTCCAGGC?TGAGAAACAG
GCCGTTCTGG?CGGAACTGGC?AGCGAAAGCC?AAACTGACGG?AACAAGTGTT?CAACGAAGCG
CCGGGTATCT?CCTGCAATCC?GGTACAAGGC?GCGATGTATA?GCTTCCCGCG?CGTTCAGCTG
CCGCCACGCG?CAGTAGAACG?TGCACAGGAA?CTGGGCCTGG?CTCCGGACAT?GTTCTTCTGT
CTGCGCCTGC?TGGAAGAAAC?CGGTATTTGC?GTTGTACCTG?GCAGCGGCTT?CGGTCAACGT
GAAGGTACTT?ACCACTTCCG?TATGACCATC?CTGCCTCCGC?TGGAAAAACT?GCGTCTGCTG
CTGGAAAAGC?TGTCCCGTTT?TCACGCGAAA?TTCACTCTGG?AATATTCTTA?G
2. prepare as claimed in claim 1 the method for sarcosine oxidase, it is characterized in that step 2) described in expression vector be: pRSF Duet, pET express series.
3. prepare as claimed in claim 1 the method for sarcosine oxidase, it is characterized in that step 2) described in coli strain be: BL21 (DE3), Rosetta (DE3).
4. prepare as claimed in claim 3 the method for sarcosine oxidase, it is characterized in that, described BL21 (DE3) bacterial strain is used for efficiently expressing the gene that is cloned in the expression vector that contains the phage t7 promotor.
5. prepare as claimed in claim 4 the method for sarcosine oxidase, it is characterized in that, lambda particles phage DE3 contains the T7 phage rna polymerase in the district, and this district is integrated in and forms described BL21 (DE3) on the karyomit(e) of BL21.
6. prepare as claimed in claim 1 the method for sarcosine oxidase, it is characterized in that step 4) described in broken bacterium method be broken, the ultrasonic or N,O-Diacetylmuramidase enzymolysis of high pressure.
7. development that utilizes product that the described method for preparing sarcosine oxidase of claim 1 makes to be used for the sarkosine detection kit as diagnostic reagent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012105784815A CN103013941A (en) | 2012-12-28 | 2012-12-28 | Preparation method and application of sarcosine oxidase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012105784815A CN103013941A (en) | 2012-12-28 | 2012-12-28 | Preparation method and application of sarcosine oxidase |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103013941A true CN103013941A (en) | 2013-04-03 |
Family
ID=47963056
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012105784815A Pending CN103013941A (en) | 2012-12-28 | 2012-12-28 | Preparation method and application of sarcosine oxidase |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103013941A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI656345B (en) * | 2016-01-13 | 2019-04-11 | Prostate cancer detection module and its operation method | |
| CN109897805A (en) * | 2019-04-04 | 2019-06-18 | 大连大学 | A kind of screening technique producing sarcosine oxidase bacterial strain |
| CN109957538A (en) * | 2019-04-04 | 2019-07-02 | 大连大学 | A kind of genetic engineering bacterium and its preparation method and application preparing sarcosine oxidase |
| CN112522223A (en) * | 2021-02-18 | 2021-03-19 | 天津科技大学 | Genetically engineered bacterium for producing L-sarcosine and construction method and application thereof |
| US11479795B2 (en) | 2021-02-18 | 2022-10-25 | Tianjin University Of Science And Technology | Genetically engineered bacterium for sarcosine production as well as construction method and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1912594A (en) * | 2006-08-10 | 2007-02-14 | 福建省洪诚生物药业有限公司 | Chemiluminescence investigating method of creatinine in serum |
| CN101426911A (en) * | 2006-04-25 | 2009-05-06 | 东洋纺织株式会社 | Modified creatinine amide hydrolase having improved affinity for substrate, and reagent composition for determination of creatinine |
-
2012
- 2012-12-28 CN CN2012105784815A patent/CN103013941A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101426911A (en) * | 2006-04-25 | 2009-05-06 | 东洋纺织株式会社 | Modified creatinine amide hydrolase having improved affinity for substrate, and reagent composition for determination of creatinine |
| CN1912594A (en) * | 2006-08-10 | 2007-02-14 | 福建省洪诚生物药业有限公司 | Chemiluminescence investigating method of creatinine in serum |
Non-Patent Citations (1)
| Title |
|---|
| 郭康平: "Bacillus sp.BSD-8肌酐降解酶系的研究", 《中国博士学位论文全文数据库》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI656345B (en) * | 2016-01-13 | 2019-04-11 | Prostate cancer detection module and its operation method | |
| CN109897805A (en) * | 2019-04-04 | 2019-06-18 | 大连大学 | A kind of screening technique producing sarcosine oxidase bacterial strain |
| CN109957538A (en) * | 2019-04-04 | 2019-07-02 | 大连大学 | A kind of genetic engineering bacterium and its preparation method and application preparing sarcosine oxidase |
| CN112522223A (en) * | 2021-02-18 | 2021-03-19 | 天津科技大学 | Genetically engineered bacterium for producing L-sarcosine and construction method and application thereof |
| US11479795B2 (en) | 2021-02-18 | 2022-10-25 | Tianjin University Of Science And Technology | Genetically engineered bacterium for sarcosine production as well as construction method and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zeng et al. | High-throughput screening technology in industrial biotechnology | |
| Steinbock et al. | The emergence of nanopores in next-generation sequencing | |
| Cabezas et al. | How to use molecular biology tools for the study of the anaerobic digestion process? | |
| Wang et al. | Development of a fluorescence assay for highly sensitive detection of Pseudomonas aeruginosa based on an aptamer-carbon dots/graphene oxide system | |
| CN111812066A (en) | Biosensor based on CRISPR/Cas12a system, kit and application of biosensor in small molecule detection | |
| US11169152B2 (en) | Function-based probes for environmental microbiome analysis and methods of making and using the same | |
| CN103013941A (en) | Preparation method and application of sarcosine oxidase | |
| Maus et al. | Biphasic study to characterize agricultural biogas plants by high-throughput 16S rRNA gene amplicon sequencing and microscopic analysis | |
| Jin et al. | A real-time LAMP-based dual-sample microfluidic chip for rapid and simultaneous detection of multiple waterborne pathogenic bacteria from coastal waters | |
| Lebuhn et al. | Microbiology and molecular biology tools for biogas process analysis, diagnosis and control | |
| CN111073892A (en) | Aptamer for identifying grouper iridovirus infected cells and construction method and application thereof | |
| CN110643611A (en) | Aptamer, construction method thereof and application thereof in detection of grouper iridovirus | |
| JP2022174242A (en) | Method for detecting target nucleic acid | |
| Song et al. | Label-free visual detection of nucleic acids in biological samples with single-base mismatch detection capability | |
| Dun-Ming et al. | Application of aptamers in food safety | |
| CN109207634A (en) | Fluorescent quantitation reference gene and its primer special and application under clerodendron trichotomum drought stress | |
| Qin et al. | Rapid detection of Pseudomonas aeruginosa using a DNAzyme‐based sensor | |
| CN105463093A (en) | Primer, kit and amplification method for fish mitochondria whole genome amplification | |
| CN103993090A (en) | Specific nucleotides for providencia O31, O41, O42, O43 and O50 and application of specific nucleotides | |
| CN106480167B (en) | Method for exploring oil gas by using microorganisms | |
| CN111349631B (en) | Aptamer specifically binding to fin algae toxin-1 and application thereof | |
| CN104328209A (en) | Primer and kit for fast detection method of leukemia minimal residual disease WT1 gene | |
| Weitong et al. | Research and application progress of microdroplets high throughput screening methods | |
| Mannu et al. | Comparison of different DNA extraction methods from peripheral blood cells: advice from the Fondazione Italiana Linfomi Minimal Residual Disease Network | |
| CN102807988A (en) | Method for preparing aspartate aminotransferase and application of aspartate aminotransferase |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20130403 |