CN102807988A - Method for preparing aspartate aminotransferase and application of aspartate aminotransferase - Google Patents
Method for preparing aspartate aminotransferase and application of aspartate aminotransferase Download PDFInfo
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- CN102807988A CN102807988A CN2012102914431A CN201210291443A CN102807988A CN 102807988 A CN102807988 A CN 102807988A CN 2012102914431 A CN2012102914431 A CN 2012102914431A CN 201210291443 A CN201210291443 A CN 201210291443A CN 102807988 A CN102807988 A CN 102807988A
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- aspartic transaminase
- aspartate aminotransferase
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Abstract
The invention discloses a method for preparing aspartate aminotransferase and application of the aspartate aminotransferase. The method includes: modifying the coding gene sequence of humanized aspartate aminotransferase subjected to gene synthesis, and removing escherichia coli rare codons; inserting the modified aspartate aminotransferase coding gene sequence into an expression vector, and using an escherichia coli strain to transform expression; inoculating strain for transforming expression to LB (Luria-Bertani) medium for IPTG (isopropyl-beta-d-thiogalactoside) induction; and homogenizing and centrifuging obtained bacteria, and enabling the centrifuged bacteria to pass through the Ni-NTA column for purification. The method for preparing the aspartate aminotransferase is extremely simple in operation, low in preparation cost and high in finished product yield and purity; the aspartate aminotransferase has high biological activity and can be used for animal immunization and screening related antibodies by serving as antigen substances; and the method and the aspartate aminotransferase lay the foundation for establishing a badly-needed calibrator preparation platform as early as possible for domestic clinical chemistry diagnosis and developing diagnostic reagent standard substances and references.
Description
Technical field
The invention belongs to genetically engineered and protein expression field; Relate to a kind of method with biotechnology efficient production aspartic transaminase; Product can be used as antigenic substance and carries out animal immune; The screening associated antibodies is diagnosed required calibration object, reference material and manufacturing of article of reference for clinical chemistry simultaneously and is laid the foundation.
Background technology
Aspartic transaminase (aspartate aminotransferase AST); Original name millet straw transferring enzyme (glutamic oxaloacetic transaminase GOT); Be transaminase important in the body, extensively exist in each tissue in vivo that AST is the highest with the heart activity.Content is very little in the normal human serum.Under the normal circumstances, the scope from hepatocellular aspartic transaminase in the blood is 0-40U/L (unit).
When the myocardial cell sustains damage, aspartic transaminase can be discharged in the blood of human body in a large number, causes that aspartic transaminase raises in the blood; When liver cell sustained damage, the value that can detect aspartic transaminase in the blood equally raise.Glutamic-oxal(o)acetic transaminase in the liver has 2 kinds of isozymes, is present in respectively in hepatocellular plastosome (mAST) and the endochylema (sAST).When the slight hepatic cell pathology, only sAST is released into blood; And when pathology was serious, mAST also can be released into blood in succession.So serum AST activity increases with the degree of hepatocellular damage.
Whether aspartic transaminase is one important in the liver function test, can check hepatic tissue impaired.In liver function test, gpt is the impaired index of reflection liver cell, and aspartic transaminase is the standard of reflection hepatic necrosis.Detection for liver cirrhosis, hepatic fibrosis, liver cancer is very accurate.Aspartic transaminase can combine the situation that reflects liver function with gpt, all below 1.0 is being liver cirrhosis or early stage liver cirrhosis patient such as gpt/aspartic transaminase ratio.
China is vast in territory, and is populous, and reagent for clinical diagnosis market is huge, has a extensive future.With regard to domestic present situation, the diagnostic reagent industry of China still is in early days of growth, i.e. the input initial stage of research and development of products and production, and also inreal arrival of growth stage, and China's diagnostic reagent market scale is about 1/14 of world market, and bigger growing space is arranged.Develop cheap and good-quality diagnostic reagent product beyond doubt very necessity and tool significance.
At present external product normally pushes market to reagent, reference article (calibration object, quality control product), instrument as a system (closed system), on detection repeatability, accuracy, very big advantage is arranged, and has corner on the market property.Experience and strength that domestic main diagnostic reagent manufacturer does not nearly all produce corresponding quality control product, the homemade goods of approving on the market is very few.
Summary of the invention
To 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 aspartic transaminase that success ratio is high.
Be the realization above-mentioned purpose, a kind of method for preparing aspartic transaminase of the present invention, this method comprises:
1) transforms the synthetic humanization aspartic transaminase coding gene sequence of full gene, remove the intestinal bacteria rare codon;
2) improved aspartic transaminase coding gene sequence in the step 1) is inserted in the expression vector, and transform expression with coli strain;
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 spinning;
Wherein, the synthetic humanization aspartic transaminase coding gene sequence of improved full gene is in the step 1):
atggctctgc?tgcattctgg?tcgtgtactg?cctggtattg?ctgctgcatt?ccacccgggc
ctggccgcgg?cggcttctgc?acgtgcatcc?tcctggtgga?ctcatgtaga?aatgggtccg
ccagatccga?tcctgggtgt?taccgaggcg?ttcaaacgcg?ataccaattc?caagaagatg
aacctgggtg?tgggcgcgta?ccgtgacgac?aatggcaaac?cttacgtact?gccatctgtg
cgtaaagcag?aagcgcaaat?cgcggctaaa?aacctggaca?aagaatatct?gccaatcggc
ggcctggcgg?aattttgtaa?agcttctgcg?gaactggcgc?tgggtgaaaa?cagcgaagtg
ctgaaaagcg?gtcgtttcgt?taccgtacag?actatctctg?gtaccggtgc?tctgcgcatt
ggtgcttctt?ttctgcaacg?tttcttcaaa?ttctctcgtg?acgtgttcct?gccgaagccg
acgtggggca?accacacccc?gatttttcgc?gatgcgggca?tgcaactgca?aggttaccgc
tactatgatc?cgaaaacttg?cggcttcgac?tttactggcg?ccgtggaaga?catctccaag
atcccggaac?agagcgttct?gctgctgcac?gcatgtgctc?ataacccaac?tggcgtagac
ccgcgtccgg?aacagtggaa?agaaattgct?accgttgtaa?agaaacgtaa?cctgttcgct
ttcttcgaca?tggcctacca?gggcttcgcg?tctggtgacg?gtgataagga?cgcgtgggcg
gttcgccact?tcattgaaca?aggtatcaat?gtatgtctgt?gccagagcta?cgccaaaaac
atgggcctgt?acggcgaacg?tgtgggcgcc?ttcaccatgg?tttgcaaaga?tgctgatgag
gctaaacgtg?tggaatctca?gctgaaaatt?ctgatccgcc?cgatgtacag?caacccacct
ctgaacggcg?ctcgtattgc?ggctgccatt?ctgaacaccc?cggacctgcg?taaacagtgg
ctgcaggagg?taaaagttat?ggctgatcgt?atcattggta?tgcgtactca?gctggtttct
aacctgaaga?aggaaggctc?cacccataac?tggcagcata?tcaccgatca?gatcggcatg
ttctgtttca?cgggtctgaa?accggagcag?gtggagcgtc?tgatcaaaga?gtttagcatt
tatatgacca?aggatggccg?tatcagcgta?gcaggtgtta?cctccagcaa?cgttggctat
ctggctcatg?ctatccatca?agtaaccaag?tag。
Further, expression vector is step 2): pRSF Duet, pET express series.
Further, coli strain is step 2): BL21 (DE3), Rosetta (DE3).
Further, said BL21 (DE3) bacterial strain is used to efficiently express the gene of cloning 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 said BL21 (DE3) on the karyomit(e) of BL21.
Further, broken bacterium method is broken, the ultrasonic or N,O-Diacetylmuramidase enzymolysis of high pressure described in the step 4).
A kind of application of method in calibration object, reference material and the preparation of reference article for preparing aspartic transaminase according to claim 1.
Further, the pure article of said preparation aspartic transaminase are added into people source or zoogenous serum, are mixed with the standard substance or the calibration object of prescribed concentration.
A kind of antigenic substance that utilizes product that the said method for preparing aspartic transaminase of claim 1 processes as immunity usefulness screens associated antibodies.
Beneficial effect of the present invention is: it is very simple that the present invention prepares the aspartic transaminase operation; Preparation cost is low, and finished product yield is high, and purity is high and have a high biological activity; Can be used as antigenic substance and carry out animal immune; The screening associated antibodies, for the calibration object of setting up domestic clinical chemistry diagnosis urgent need as early as possible prepares platform, development diagnostic reagent reference material and reference article lay the foundation.
Description of drawings
Fig. 1 obtains the insertion fragment of about 1.5kb for the 6His-AST pRSF that makes up cuts with BamH I and EcoR I enzyme, shows that goal gene has inserted carrier, makes up successfully;
Fig. 2 is the SDS-PAGE electrophorogram of the AST finished product of purifying, and product purity can reach more than 90%.
Embodiment
Below, with reference to accompanying drawing, the present invention is more comprehensively explained, exemplary embodiment of the present invention has been shown in the accompanying drawing.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, and scope of the present invention is fully conveyed to those of ordinary skill in the art comprehensively with complete.
The present invention has set up a kind of method for preparing aspartic transaminase simple to operate, rapid, lower-cost.The principle of this method mainly is: the aspartic transaminase encoding sox after will optimizing inserts the expression vector that builds, and in intestinal bacteria, induces it to efficiently express, and this method is specially:
1. the structure of aspartic transaminase expression vector:
Transform humanization aspartic transaminase coding gene sequence, remove the intestinal bacteria rare codon, the synthetic improved AST encoding sequence of the full gene of invitrogen company from Beijing;
Utilize http://www.faculty.ucr.edu/ ~ mmaduro/codonusage/usage.htm web page analysis, in original AST coding gene sequence, codon usage frequency accounts for 16% 10% with interior in the intestinal bacteria; Improved gene order is removed the intestinal bacteria rare codon fully, and e. coli codon is activated fully.
Utilize BamH I and EcoRI site to be cloned in the PRSF Duet carrier through genetic engineering means; And transformed into escherichia coli TOP10 bacterial strain; Several transformed clones of picking extract plasmid, identify through digestion with restriction enzyme whether goal gene inserts carrier (as shown in Figure 1), choose the plasmid that inserts goal gene and carry out determined dna sequence; The result shows that the aspartic transaminase gene successfully is cloned in the pRSF Duet carrier, and we are with its called after 6His-AST pRSF.
The synthetic humanization aspartic transaminase coding gene sequence of improved full gene is:
atggctctgc?tgcattctgg?tcgtgtactg?cctggtattg?ctgctgcatt?ccacccgggc
ctggccgcgg?cggcttctgc?acgtgcatcc?tcctggtgga?ctcatgtaga?aatgggtccg
ccagatccga?tcctgggtgt?taccgaggcg?ttcaaacgcg?ataccaattc?caagaagatg
aacctgggtg?tgggcgcgta?ccgtgacgac?aatggcaaac?cttacgtact?gccatctgtg
cgtaaagcag?aagcgcaaat?cgcggctaaa?aacctggaca?aagaatatct?gccaatcggc
ggcctggcgg?aattttgtaa?agcttctgcg?gaactggcgc?tgggtgaaaa?cagcgaagtg
ctgaaaagcg?gtcgtttcgt?taccgtacag?actatctctg?gtaccggtgc?tctgcgcatt
ggtgcttctt?ttctgcaacg?tttcttcaaa?ttctctcgtg?acgtgttcct?gccgaagccg
acgtggggca?accacacccc?gatttttcgc?gatgcgggca?tgcaactgca?aggttaccgc
tactatgatc?cgaaaacttg?cggcttcgac?tttactggcg?ccgtggaaga?catctccaag
atcccggaac?agagcgttct?gctgctgcac?gcatgtgctc?ataacccaac?tggcgtagac
ccgcgtccgg?aacagtggaa?agaaattgct?accgttgtaa?agaaacgtaa?cctgttcgct
ttcttcgaca?tggcctacca?gggcttcgcg?tctggtgacg?gtgataagga?cgcgtgggcg
gttcgccact?tcattgaaca?aggtatcaat?gtatgtctgt?gccagagcta?cgccaaaaac
atgggcctgt?acggcgaacg?tgtgggcgcc?ttcaccatgg?tttgcaaaga?tgctgatgag
gctaaacgtg?tggaatctca?gctgaaaatt?ctgatccgcc?cgatgtacag?caacccacct
ctgaacggcg?ctcgtattgc?ggctgccatt?ctgaacaccc?cggacctgcg?taaacagtgg
ctgcaggagg?taaaagttat?ggctgatcgt?atcattggta?tgcgtactca?gctggtttct
aacctgaaga?aggaaggctc?cacccataac?tggcagcata?tcaccgatca?gatcggcatg
ttctgtttca?cgggtctgaa?accggagcag?gtggagcgtc?tgatcaaaga?gtttagcatt
tatatgacca?aggatggccg?tatcagcgta?gcaggtgtta?cctccagcaa?cgttggctat
ctggctcatg?ctatccatca?agtaaccaag?tag。
2. the expression of aspartic transaminase:
1) is the host bacterium with BL21 (DE3), 6His-AST pRSF transformed that picking list bacterium colony spends the night in 3ml LB culture medium culturing;
2) with the 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 to efficiently express the gene of cloning in the expression vector that contains the phage t7 promotor (like pET series).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 aspartic transaminase:
1) get 400ml nutrient solution 3000rpm spinning 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, and 12,000rpm, 4 ℃ of centrifugal 10min, supernatant and Ni-NTA were hatched 1 hour on ice;
2) cross the Ni-NTA affinity column,, use the 250mMImidazle wash-out at last, collect elutriant with protein extraction buffer washing.
3) the albumen elutriant is crossed the G50 post and is removed Imidazle, can obtain the AST (as shown in Figure 2) of 90% above purity like this.
4. aspartic transaminase determination of activity:
The pure article of preparation aspartic transaminase are measured active with Olympus AU5400 biochemical instruments, enzyme work can reach 30000U/L, and frozen 6 months of-20 ℃ of glycerine, and enzyme work remains unchanged basically.These pure article can be added into people source or zoogenous serum, are mixed with the standard substance or the calibration object of prescribed concentration.
The above-mentioned method for preparing aspartic transaminase can be used as antigenic substance and carries out animal immune, the screening associated antibodies, and in calibration object, reference material and the preparation of reference article, use, not to be diagnosed as purpose, not belong to the diagnostic method of disease.
Only if 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 exchange with its full name and use.
Only if special indicating, the present invention uses but clearly sets forth or simple technology and the method for setting forth is meant normally used technology in present technique field 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.
SEQUENCE?LISTING
< 110>the lucky sci-tech consultation of Beijing Xiang Dasheng ltd
< 120>a kind of method and application for preparing aspartic transaminase
< 130>invention
<160> 1
<170> PatentIn?version?3.3
<210> 1
<211> 24
<212> DNA
<213> AST?Gene?sequence
<400> 1
atggctctgc?tgcattctgg?tcgtgtactg?cctggtattg?ctgctgcatt?ccacccgggc?ctggccgcgg?cggcttctgc?acgtgcatcc?tcctggtgga?ctcatgtaga?aatgggtccg?ccagatccga?tcctgggtgt?taccgaggcg?ttcaaacgcg?ataccaattc?caagaagatg?aacctgggtg?tgggcgcgta?ccgtgacgac?aatggcaaac?cttacgtact?gccatctgtg?cgtaaagcag?aagcgcaaat?cgcggctaaa?aacctggaca?aagaatatct?gccaatcggc?ggcctggcgg?aattttgtaa?agcttctgcg?gaactggcgc?tgggtgaaaa?cagcgaagtg?ctgaaaagcg?gtcgtttcgt?taccgtacag?actatctctg?gtaccggtgc?tctgcgcatt?ggtgcttctt?ttctgcaacg?tttcttcaaa?ttctctcgtg?acgtgttcct?gccgaagccg?acgtggggca?accacacccc?gatttttcgc?gatgcgggca?tgcaactgca?aggttaccgc?tactatgatc?cgaaaacttg?cggcttcgac?tttactggcg?ccgtggaaga?catctccaag?atcccggaac?agagcgttct?gctgctgcac?gcatgtgctc?ataacccaac?tggcgtagac?ccgcgtccgg?aacagtggaa?agaaattgct?accgttgtaa?agaaacgtaa?cctgttcgct?ttcttcgaca?tggcctacca?gggcttcgcg?tctggtgacg?gtgataagga?cgcgtgggcg?gttcgccact?tcattgaaca?aggtatcaat?gtatgtctgt?gccagagcta?cgccaaaaac?atgggcctgt?acggcgaacg?tgtgggcgcc?ttcaccatgg?tttgcaaaga?tgctgatgag?gctaaacgtg?tggaatctca?gctgaaaatt?ctgatccgcc?cgatgtacag?caacccacct?ctgaacggcg?ctcgtattgc?ggctgccatt?ctgaacaccc?cggacctgcg?taaacagtgg?ctgcaggagg?taaaagttat?ggctgatcgt?atcattggta?tgcgtactca?gctggtttct?aacctgaaga?aggaaggctc?cacccataac?tggcagcata?tcaccgatca?gatcggcatg?ttctgtttca?cgggtctgaa?accggagcag?gtggagcgtc?tgatcaaaga?gtttagcatt?tatatgacca?aggatggccg?tatcagcgta?gcaggtgtta?cctccagcaa?cgttggctat?ctggctcatg?ctatccatca?agtaaccaag?tag 1293
Claims (9)
1. a method for preparing aspartic transaminase is characterized in that, this method comprises:
1) transforms the synthetic humanization aspartic transaminase coding gene sequence of full gene, remove the intestinal bacteria rare codon;
2) improved aspartic transaminase coding gene sequence in the step 1) is inserted in the expression vector, and transform expression with coli strain;
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 spinning;
Wherein, the synthetic humanization aspartic transaminase coding gene sequence of improved full gene is in the step 1):
atggctctgc?tgcattctgg?tcgtgtactg?cctggtattg?ctgctgcatt?ccacccgggc
ctggccgcgg?cggcttctgc?acgtgcatcc?tcctggtgga?ctcatgtaga?aatgggtccg
ccagatccga?tcctgggtgt?taccgaggcg?ttcaaacgcg?ataccaattc?caagaagatg
aacctgggtg?tgggcgcgta?ccgtgacgac?aatggcaaac?cttacgtact?gccatctgtg
cgtaaagcag?aagcgcaaat?cgcggctaaa?aacctggaca?aagaatatct?gccaatcggc
ggcctggcgg?aattttgtaa?agcttctgcg?gaactggcgc?tgggtgaaaa?cagcgaagtg
ctgaaaagcg?gtcgtttcgt?taccgtacag?actatctctg?gtaccggtgc?tctgcgcatt
ggtgcttctt?ttctgcaacg?tttcttcaaa?ttctctcgtg?acgtgttcct?gccgaagccg
acgtggggca?accacacccc?gatttttcgc?gatgcgggca?tgcaactgca?aggttaccgc
tactatgatc?cgaaaacttg?cggcttcgac?tttactggcg?ccgtggaaga?catctccaag
atcccggaac?agagcgttct?gctgctgcac?gcatgtgctc?ataacccaac?tggcgtagac
ccgcgtccgg?aacagtggaa?agaaattgct?accgttgtaa?agaaacgtaa?cctgttcgct
ttcttcgaca?tggcctacca?gggcttcgcg?tctggtgacg?gtgataagga?cgcgtgggcg
gttcgccact?tcattgaaca?aggtatcaat?gtatgtctgt?gccagagcta?cgccaaaaac
atgggcctgt?acggcgaacg?tgtgggcgcc?ttcaccatgg?tttgcaaaga?tgctgatgag
gctaaacgtg?tggaatctca?gctgaaaatt?ctgatccgcc?cgatgtacag?caacccacct
ctgaacggcg?ctcgtattgc?ggctgccatt?ctgaacaccc?cggacctgcg?taaacagtgg
ctgcaggagg?taaaagttat?ggctgatcgt?atcattggta?tgcgtactca?gctggtttct
aacctgaaga?aggaaggctc?cacccataac?tggcagcata?tcaccgatca?gatcggcatg
ttctgtttca?cgggtctgaa?accggagcag?gtggagcgtc?tgatcaaaga?gtttagcatt
tatatgacca?aggatggccg?tatcagcgta?gcaggtgtta?cctccagcaa?cgttggctat
ctggctcatg?ctatccatca?agtaaccaag?tag
2. prepare the method for aspartic transaminase according to claim 1, it is characterized in that step
2) expression vector described in is: pRSF Duet, pET express series.
3. prepare the method for aspartic transaminase according to claim 1, it is characterized in that step 2) described in coli strain be: BL21 (DE3), Rosetta (DE3).
4. like the said method for preparing aspartic transaminase of claim 3, it is characterized in that said BL21 (DE3) bacterial strain is used to efficiently express the gene of cloning in the expression vector that contains the phage t7 promotor.
5. like the said method for preparing aspartic transaminase of claim 4, it is characterized in that lambda particles phage DE3 contains the T7 phage rna polymerase in the district, this district is integrated in and forms said BL21 (DE3) on the karyomit(e) of BL21.
6. prepare the method for aspartic transaminase according to claim 1, it is characterized in that, broken bacterium method is broken, the ultrasonic or N,O-Diacetylmuramidase enzymolysis of high pressure described in the step 4).
7. the application of method in calibration object, reference material and the preparation of reference article for preparing aspartic transaminase according to claim 1.
8. application as claimed in claim 7 is characterized in that, the pure article of said preparation aspartic transaminase are added into people source or zoogenous serum, is mixed with the standard substance or the calibration object of prescribed concentration.
9. an antigenic substance that utilizes product that the said method for preparing aspartic transaminase of claim 1 processes as immunity usefulness screens associated antibodies.
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| CN2012102914431A CN102807988A (en) | 2012-08-15 | 2012-08-15 | Method for preparing aspartate aminotransferase and application of aspartate aminotransferase |
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| CN2012102914431A CN102807988A (en) | 2012-08-15 | 2012-08-15 | Method for preparing aspartate aminotransferase and application of aspartate aminotransferase |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104447998A (en) * | 2014-12-22 | 2015-03-25 | 宁波美康盛德医学检验所有限公司 | Method for preparing polyclonal antibody of human aspartic transaminase cytoplasm isozymes |
| RU2703169C1 (en) * | 2018-06-20 | 2019-10-15 | Общество с ограниченной ответственностью научно-технический центр "БиоКлиникум" (ООО НТЦ "БиоКлиникум") | Strain-producer of human glutamate oxaloacetate transaminase |
-
2012
- 2012-08-15 CN CN2012102914431A patent/CN102807988A/en active Pending
Non-Patent Citations (4)
| Title |
|---|
| NCBI: "aspartate aminotransferase,mitochondrial precursor[Homo sapiens]", 《REFERENCE SEQUENCE: NP_002071.2》 * |
| 宫长斌 等: "组成型天冬氨酸转氨酶基因工程菌的构建和高效表达", 《过程工程学报》 * |
| 廖红梅 等: "人丙氨酸氨基转移酶基因（ALT1）的克隆、表达、纯化及酶活性的鉴定", 《沈阳药科大学学报》 * |
| 李霜 等: "天冬氨酸转氨酶在大肠杆菌中分泌表达", 《第一届全国化学工程与生物化工年会论文摘要集（下）》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104447998A (en) * | 2014-12-22 | 2015-03-25 | 宁波美康盛德医学检验所有限公司 | Method for preparing polyclonal antibody of human aspartic transaminase cytoplasm isozymes |
| RU2703169C1 (en) * | 2018-06-20 | 2019-10-15 | Общество с ограниченной ответственностью научно-технический центр "БиоКлиникум" (ООО НТЦ "БиоКлиникум") | Strain-producer of human glutamate oxaloacetate transaminase |
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