CN108610406A - A kind of 3 genes of oyster cell apoptogene caspase and its application in preparing pathological examination diagnostic reagent - Google Patents
A kind of 3 genes of oyster cell apoptogene caspase and its application in preparing pathological examination diagnostic reagent Download PDFInfo
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Abstract
The present invention discloses a kind of 3 genes of oyster cell apoptogene caspase and its application in preparing pathological examination diagnostic reagent.The inventors discovered that significant 3 genes of gene caspase of the huge oyster cell apoptosis in a Hong Kong, its nucleotide sequence is as shown in SEQ ID NO.1, it is significantly raised after vibrio alginolyticus (Vibrio alginolyticus), staphylococcus (Staphylococcus haemolyticus) infection, it can be as the standard of detection oyster cell apoptosis situation, a positive and accurate restricted speed signal is provided for the prevention of oyster disease, it is therefore prevented that the expansion of disaster.The application of the technology, by the further germplasm innovation ability for promoting cultivated shellfish, the High-efficiency Sustainable sexual development of supporting industry.
Description
Technical field：
The present invention relates to the immune molecule mechanisms with Apoptosis of shellfish, are more particularly to a kind of from the post-stimulatory perfume (or spice) of vibrios
3 bases of caspase screened in the huge oyster in port (Crassostrea hongkongensis) haemocyte subtractive Hybrid Library
The purposes of the nucleotide sequence of cause, amino acid sequence and preparation method thereof and the gene.
Background technology：
China's oyster annual output accounts for 80% of Gross World Product or more, is maximum oyster culture state in the world.Hong Kong is male
Oyster is widely distributed in the coastal marine site in the place such as Guangxi to Fujian, is the most important oyster culture kind of the South China coastal.Due to
Its breeding way is based on tradition cultivation, and the limitations such as wide, high energy consumption that there is cultured areas, yield is also because various conditions become
Change and fluctuates.As marine environmental pollution is increasingly severe, the risk of oyster culture is also increasing.China is at present about oyster
The technology of Defect inspection falls behind very much, substantially belongs to space state, therefore, improves oyster health monitoring technique, takes precautions against disease in time
Harmful propagation is development and raising oyster culture industry effective means.
Apoptosis is a kind of fundamental mechanism that cell deletion is carried out in multicellular organism normal development, atomization, with
Tissue is closely related from steady, aging and cellular damage.When mechanism is by disease, environment-stress, apoptotic pathways will
Start, a series of protease will be activated, and make cell is orderly to be degraded.In this above process, there is class of enzymes to exist
Serve during this key, their structure feature and cysteine proteinase are much like, can specifically cut
The peptide bond after Asp is cut, therefore is named as caspase.It is that Apoptosis closes that, which there are many members, wherein caspase 3 in Caspase families,
The execution enzyme of key, the typically significant gene of Apoptosis whether carried out have important answer in biological pathology detection
With value.
Invention content：
The first purpose of the invention is to provide the relevant gene order-caspase of a kind of Hong Kong oyster Apoptosis 3
Gene.
Relevant 3 genes of gene order-caspase of Hong Kong oyster Apoptosis of the present invention, nucleotide sequence is such as
Shown in SEQ ID NO.1,296 amino acid are encoded, amino acid sequence is as shown in SEQ ID NO.2.
The present invention is on the basis of the huge oyster transcript profile sequencing in Hong Kong, it was found that the significant gene of the Species Cell apoptosis
3 genes of caspase.By RACE technologies, we obtain the overall length of the gene, nucleotide sequence such as SEQ ID NO.1 institutes
Show, the amino acid sequence of the gene code is as shown in SEQ ID NO.2.Quantitative PCR experiment shows 3 genes of caspase molten
After algae vibrios (Vibrio alginolyticus), staphylococcus (Staphylococcus haemolyticus) infection significantly
Up-regulation, can be positive and accurate for the prevention offer one of oyster disease as the standard of detection oyster cell apoptosis situation
Restricted speed signal, it is therefore prevented that the expansion of disaster.
Therefore, second object of the present invention is to provide the detection reagents of expression quantity of detection 3 genes of caspase and is making
Application in standby oyster disease prevention warning reagent.
The oyster disease prevention warning reagent is the reagent for detecting oyster cell apoptosis situation.
The oyster is the huge oyster in Hong Kong.
The reagent of the detection oyster cell apoptosis situation is preferably due to vibrio alginolyticus and/or staphy lococcus infection is male
The detection reagent of oyster cell apoptosis situation is detected after oyster.
Third object of the present invention is to provide a kind of detection primers of 3 genes of above-mentioned caspase；
For 3 genes of caspase：
Chcaspase3-F:5'-GGAGCGGTTTCAGGACTTGG-3'
Chcaspase3-R:5'-AGCATTCCGTCGGTAGCGTA-3'。
The detection primer that fourth object of the present invention is to provide 3 genes of above-mentioned caspase is preparing detection caspase
Application in the expression quantity reagent of 3 genes.
It is preferred that further including the detection primer for having reference gene GAPDH, specially：
ChGAPDH-F:5'-GGATTGGCGTGGTGGTAGAG-3'
ChGAPDH-R:5'-GTATGATGCCCCTTTGTTGAGTC-3'。
Fifth object of the present invention is to provide a kind of quantitative detecting methods of 3 genes of caspase, which is characterized in that is
Using the reverse transcription product of oyster total serum IgE as template, using the detection primer of 3 genes of above-mentioned caspase as primer, quantitative PCR is utilized
Detect the expression quantity of 3 genes of caspase.
It is preferred that also using gene GAPDH as internal reference.
It is preferred that the reaction system of the quantitative PCR is：
Response procedures are：95 DEG C of 5min, 1 cycle；95 DEG C of 10s, 58.5 DEG C of 10s, 72 DEG C of 20s, 45 cycles.
Sixth object of the present invention is to provide 3 genes of above-mentioned caspase to exist as the significant gene of oyster cell apoptosis
Prepare the application in the reagent of detection oyster cell apoptosis situation.
The inventors discovered that significant 3 genes of gene-caspase of the huge oyster cell apoptosis in a Hong Kong, molten
After algae vibrios (Vibrio alginolyticus), staphylococcus (Staphylococcus haemolyticus) infection significantly
Up-regulation, can be positive and accurate for the prevention offer one of oyster disease as the standard of detection oyster cell apoptosis situation
Restricted speed signal, it is therefore prevented that the expansion of disaster.The application of the technology will further promote the germplasm innovation ability of cultivated shellfish,
The High-efficiency Sustainable sexual development of supporting industry.
Description of the drawings：
Fig. 1 is the Tissue distribution of 3 albumen of caspase, real-time quantitative PCR the experimental results showed that 3 albumen of caspase each
There is expression in a tissue, content is minimum in heart, and expression quantity highest in haemocyte.Different lowercase letter sample rooms
Significant difference.
Fig. 2 is the expression of vibrio alginolyticus stimulation induction 3 genes of caspase.After pathogen stimulation, caspase in haemocyte
The expression of 3 genes is significantly raised.Asterisk (* *) indicates and control group difference is extremely notable.
Fig. 3 is the expression of staphylococcus stimulation induction 3 genes of caspase.After pathogen stimulation, caspase in haemocyte
The expression of 3 genes is significantly raised.Asterisk (* *) indicates and control group difference is extremely notable.
Specific implementation mode：
Implement to be the further explanation to the present invention below, rather than limiting the invention.
Embodiment 1：
1.RNA is extracted and reverse transcription
A) it after Hong Kong oysters cell or tissue adds Trizol, is ground using tissue grinder, is placed at room temperature for 5min, make it
Fully cracking.
B) .12,000rpm centrifuge 5min, abandon precipitation.
C) chloroform is added by 200ul chloroforms/ml Trizol in, and 15min is placed at room temperature for after vibrating mixing.
D) .4 DEG C of 12,000g centrifuges 15min.
E) draws upper strata aqueous phase, until in another centrifuge tube.
F) isopropanol mixing is added by 0.5ml isopropanols/ml Trizol in, is placed at room temperature for 5-10min.
G) .4 DEG C 12,000g centrifuge 10min, abandon supernatant, RNA is sunken to tube bottom.
H) 75% ethyl alcohol is added by 75% ethyl alcohol of 1ml/ml Trizol in, mildly vibrates centrifuge tube, and suspend precipitation.
I) .4 DEG C 8,000g centrifuge 5min, abandon supernatant as possible.
J) 5-10min is dried or be dried in vacuo to room temperatures.
K) can use 50ul H2O, TE buffer or 0.5%SDS to dissolve RNA sample, 55-60 DEG C, 5-10min.
L) is after being incubated at room temperature 15min, and 250 μ LDNase Stop Solution (DSA) are added, 13,000 × g
Centrifuge 1min.
M) repeats step a-j, adds 100 μ L nuclease-free waters, and then, 13,000 × g centrifuges 1min, abandons centrifugal column, makes
The concentration of RNA in the solution collected by spectrophotometer measurement after packing, is stored in -80 DEG C.
N) synthesis of first chain cDNA of
1) DNase I digest
The RNA of each group is added in PCR centrifuge tubes respectively according to following component
| RNA | 1μg |
| 10XDNase I buffer | 1.2μL |
| DNase I | 1μL |
| Sterile H2O | Up to 12μL |
2) it gently blows and beats mixing with pipette tips and slightly centrifuges；37 DEG C, 15min, digest the DNA in RNA
3) 1 μ L EDTA mixings are added, 65 DEG C, 10min makes DNase I inactivate
4) it is sequentially added centrifuge tube according to following component：
Mixing is gently blown and beaten with pipette tips and is slightly centrifuged, and is positioned over 37 DEG C, 30min；
5) 85 DEG C, 5s makes reverse transcriptase inactivate, then 4 DEG C of short-term preservations, -20 DEG C of long-term preservations.
2, subtractive Hybrid Library is built
Subtractive Hybrid Library using PCR-Select cDNA Subtraction Kit (Clontech, USA) kits into
Row structure.After the huge oyster in Hong Kong is stimulated 8h by vibrios, haemocyte is collected, extracts haemocyte total serum IgE according to above-mentioned method, and make
Haemocyte total serum IgE (is not handled) by vibrios with same method extraction control group.The huge oyster in Hong Kong is as tester using after stimulation,
Control group is deducted hybridization as driver.CDNAs after subtractive is hybridized is subcloned into pGEM-Teasy vector
(Promega, USA) and it is transformed into Escherichia coli JM109 (Promega, USA) competent cell.It selects at random
2000 are cloned and are sequenced.
The clone of 3 full length genes of 3.caspase
According to the obtained cloning and sequencing of step 2, is analyzed by biosoftwares such as BLASTEN and determine 3 sequences of caspase
Row, then using this sequence as masterplate, carried out by GeneRacerTM RACE Ready cDNA kits (Invitrogen, USA)
5'/3'RACE of the gene, concrete operations are with reference to specification.After obtaining RACE sequences, by splicing, the overall length of the gene is obtained
CDNA analyzes its code area, and the nucleotide sequence of code area is named as 3 genes of caspase as shown in SEQ ID NO.1,
The amino acid sequence that it is encoded is named as 3 albumen of caspase as shown in SEQ ID NO.2.
4. real-time quantitative PCR
It is LightCycler 480SYBR Green I (Roch) to carry out kit used in real-time quantitative PCR, is made
Instrument is LightCycler 480System (Roche), and used template is the reverse transcription product of total serum IgE, is used
Internal reference GAPDH, used primer has：
For 3 genes of caspase：Chcaspase3-F:5'-GGAGCGGTTTCAGGACTTGG-3'
Chcaspase3-R:5'-AGCATTCCGTCGGTAGCGTA-3'
For internal reference GAPDH genes：ChGAPDH-F:5'-GGATTGGCGTGGTGGTAGAG-3'
ChGAPDH-R:5'-GTATGATGCCCCTTTGTTGAGTC-3'
QRT-PCR experiments are as follows using Light-Cycler 480II System (Roche) progress reaction system：
Thermal cycle conditions are as follows：95 DEG C of 5min, 1 cycle；95 DEG C of 10s, 58.5 DEG C of 10s, 72 DEG C of 20s, 45 cycles.Make
With 2Δ Δ CTMethod carries out relative quantification to transcript.Experimental group and control group are respectively provided with 3 parallel reactions.
5. Tissue distribution and sample treatment
The total serum IgE for extracting each tissue of 5 huge oysters in Hong Kong respectively (is cDNA see step 1) and reverse transcription, uses
Real-time round pcrs have detected 3 genes of caspase in the lymphocyte of the huge oyster in Hong Kong, and the cheek touches lip, sexual gland, digestion
Gland, outer embrane, cardiocoelom, the distribution situation in closed shell flesh.As a result its wide expression, wherein outer embrane in each tissue are shown
Middle expression quantity is minimum, in hemolymph expression highest (Fig. 1).
The huge oyster in normal Hong Kong is taken, injects 1.0 × 10 respectively9The pathogen of a work, using PBS injections group as reference, not
Same time point grab sample (5 one group), it is cDNA to extract the total serum IgE of haemocyte and reverse transcription, sees step 1.As mould
Version carries out the expression quantity of Real-time PCR detection 3 genes of caspase of sample, as a result, it has been found that vibrio alginolyticus (Fig. 2), Portugal
Grape coccus (Fig. 3) infects the notable up-regulated expression that can cause 3 genes of caspase in haemocyte.
Sequence table
<110>Chinese Academy of Science Nanhai Ocean Research Institute
<120>A kind of 3 genes of oyster cell apoptogene caspase and its application in preparing pathological examination diagnostic reagent
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 891
<212> DNA
<213>The huge oyster in Hong Kong (Crassostrea hongkongensis)
<400> 1
atgtcgtccc cccggcgtag tccggtctcc gtcgaccacg gatctcccta cttactgagg 60
aaagccagcg gagaccgcag atcggttcgt ccaggtgccc catcggagcc aaatgagtac 120
aactttaact atcacaaaag aggccttttt atcattatca ataacaagaa tttccatccg 180
tccaccggca aagagtcacg cgaagggact gacgtagacg cggagagact ggaggagcgg 240
tttcaggact tggggttcga tgttcgacgc tacaacgacg tgtcgtcatc caaactattg 300
caacttatga acgaagcctc acagcttgac cattccgatt ccgactgttt cggttgtgcc 360
attctgagtt atggaattga ggggagggtc tacgctaccg acggaatgct gcctttagac 420
gtcctcatcg ttcccttcaa gggagacaag tgcccgatgt tggttggcaa acccaaactt 480
ttttttctac agtcttgtcg ctgtccaaac ttagaacagt ctgttgaaga ctcggtgtcc 540
tataaaagct tcctgtccga ggattccgag cggagtttct ccgggatgag gaggatccca 600
gttgaggctg acttcttgtt cttctattcc acagtacctg gtttctattc atggcggaat 660
caccaggaag gatcctggct tatccaagcc ttgtgtatcg ttttggagaa ctatggttcc 720
aaaatggaac ttctgcacat gctcactcaa gtcaatcgca tggcggcgta tgaattcgag 780
tcgtgttcgg atgaacattt tacggatgag gtcaaacaga tgccttgtat tgtgtccatg 840
cttaccagat atgtgtactt ccggcctaag aaaccggata tccgggacta a 891
<210> 2
<211> 296
<212> PRT
<213>The huge oyster in Hong Kong (Crassostrea hongkongensis)
<400> 2
Met Ser Ser Pro Arg Arg Ser Pro Val Ser Val Asp His Gly Ser Pro
1 5 10 15
Tyr Leu Leu Arg Lys Ala Ser Gly Asp Arg Arg Ser Val Arg Pro Gly
20 25 30
Ala Pro Ser Glu Pro Asn Glu Tyr Asn Phe Asn Tyr His Lys Arg Gly
35 40 45
Leu Phe Ile Ile Ile Asn Asn Lys Asn Phe His Pro Ser Thr Gly Lys
50 55 60
Glu Ser Arg Glu Gly Thr Asp Val Asp Ala Glu Arg Leu Glu Glu Arg
65 70 75 80
Phe Gln Asp Leu Gly Phe Asp Val Arg Arg Tyr Asn Asp Val Ser Ser
85 90 95
Ser Lys Leu Leu Gln Leu Met Asn Glu Ala Ser Gln Leu Asp His Ser
100 105 110
Asp Ser Asp Cys Phe Gly Cys Ala Ile Leu Ser Tyr Gly Ile Glu Gly
115 120 125
Arg Val Tyr Ala Thr Asp Gly Met Leu Pro Leu Asp Val Leu Ile Val
130 135 140
Pro Phe Lys Gly Asp Lys Cys Pro Met Leu Val Gly Lys Pro Lys Leu
145 150 155 160
Phe Phe Leu Gln Ser Cys Arg Cys Pro Asn Leu Glu Gln Ser Val Glu
165 170 175
Asp Ser Val Ser Tyr Lys Ser Phe Leu Ser Glu Asp Ser Glu Arg Ser
180 185 190
Phe Ser Gly Met Arg Arg Ile Pro Val Glu Ala Asp Phe Leu Phe Phe
195 200 205
Tyr Ser Thr Val Pro Gly Phe Tyr Ser Trp Arg Asn His Gln Glu Gly
210 215 220
Ser Trp Leu Ile Gln Ala Leu Cys Ile Val Leu Glu Asn Tyr Gly Ser
225 230 235 240
Lys Met Glu Leu Leu His Met Leu Thr Gln Val Asn Arg Met Ala Ala
245 250 255
Tyr Glu Phe Glu Ser Cys Ser Asp Glu His Phe Thr Asp Glu Val Lys
260 265 270
Gln Met Pro Cys Ile Val Ser Met Leu Thr Arg Tyr Val Tyr Phe Arg
275 280 285
Pro Lys Lys Pro Asp Ile Arg Asp
290 295
Claims (10)
- 3 albumen of 1.caspase, which is characterized in that amino acid sequence is as shown in SEQ ID NO.2.
- 2. the encoding gene of coding 3 albumen of caspase described in claim 1.
- 3. encoding gene according to claim 2, which is characterized in that it is 3 genes of caspase, and nucleotide sequence is such as Shown in SEQ ID NO.1.
- 4. a kind of test right requires the detection reagent of the expression quantity of 3 genes of caspase described in 3 pre- in preparation oyster disease Application in anti-warning reagent.
- 5. application according to claim 4, which is characterized in that the oyster disease prevention warning reagent is detection oyster The reagent of Apoptosis situation.
- 6. application according to claim 4 or 5, which is characterized in that the oyster is the huge oyster in Hong Kong.
- 7. application according to claim 5, which is characterized in that the reagent of the detection oyster cell apoptosis situation be by The detection reagent of oyster cell apoptosis situation is detected after vibrio alginolyticus and/or staphy lococcus infection oyster.
- 8. a kind of detection primer of 3 genes of caspase described in claim 3；The detection primer of 3 genes of caspase is：Chcaspase3-F:5'-GGAGCGGTTTCAGGACTTGG-3'Chcaspase3-R:5'-AGCATTCCGTCGGTAGCGTA-3'。
- 9. the detection primer of 3 genes of caspase according to any one of claims 8 is in the expression quantity examination for preparing detection 3 genes of caspase Application in agent.
- 10. 3 genes of caspase described in claim 3 are thin in preparation detection oyster as the significant gene of oyster cell apoptosis Application in the reagent of born of the same parents' apoptosis situation.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112300259A (en) * | 2020-11-06 | 2021-02-02 | 中国科学院南海海洋研究所 | Autophagy protein beclin-1 and coding gene and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2016065048A (en) * | 2014-09-17 | 2016-04-28 | 御木本製薬株式会社 | Caspase-14 expression accelerator |
| CN107540738A (en) * | 2017-04-10 | 2018-01-05 | 清远职业技术学院 | A kind of huge oyster Galectins ChGalectin in Hong Kong and its preparation method and application |
-
2018
- 2018-04-04 CN CN201810301750.0A patent/CN108610406B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2016065048A (en) * | 2014-09-17 | 2016-04-28 | 御木本製薬株式会社 | Caspase-14 expression accelerator |
| CN107540738A (en) * | 2017-04-10 | 2018-01-05 | 清远职业技术学院 | A kind of huge oyster Galectins ChGalectin in Hong Kong and its preparation method and application |
Non-Patent Citations (2)
| Title |
|---|
| ZHANG G等: "ACCESSION：XP_011445227：PREDICTED: caspase-3 isoform X2 [Crassostrea gigas]", 《GENBANK》 * |
| ZHIMING XIANG等: "Cloning, characterization and expression analysis of a caspase-8 like gene from the Hong Kong oyster, Crassostrea hongkongensis", 《FISH & SHELLFISH IMMUNOLOGY》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112300259A (en) * | 2020-11-06 | 2021-02-02 | 中国科学院南海海洋研究所 | Autophagy protein beclin-1 and coding gene and application thereof |
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