CN108486122A - A kind of the DNA bar code standard sequence and its method for identifying molecules of South Mountain Storehouse midge - Google Patents
A kind of the DNA bar code standard sequence and its method for identifying molecules of South Mountain Storehouse midge Download PDFInfo
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- CN108486122A CN108486122A CN201810083833.7A CN201810083833A CN108486122A CN 108486122 A CN108486122 A CN 108486122A CN 201810083833 A CN201810083833 A CN 201810083833A CN 108486122 A CN108486122 A CN 108486122A
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- midge
- storehouse midge
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6869—Methods for sequencing
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
Abstract
The present invention discloses a kind of the DNA bar code standard sequence and its method for identifying molecules of South Mountain Storehouse midge, it is characterized in that obtaining I gene orders of CO of the Storehouse midge using the method for molecular biology DNA sequencing, it is compared again by gene order similitude, to carry out Identification of Species to the Storehouse midge according to comparison result.Storehouse midge method for identifying molecules in South Mountain provided by the invention, is advantageously implemented it and fast and accurately identifies.
Description
Technical field
The present invention relates to insect species identify field, and in particular to the DNA bar code standard sequence of South Mountain Storehouse midge and its point
Sub- identification method.
Background technology
Storehouse midge（CulicoidesLatreille, 1809）It is under the jurisdiction of Insecta（Insecta）Diptera（diptera）
Heleidae（Ceratopogonidae）, build is small, type is various, widely distributed, the whole world it is 1368 kinds existing, China
Know 347 kinds, and worldwide also constantly finds new species.Blood sucking midge refer to can by piercing and sucking ruminant blood come
Propagate the midge of entomophila pathogen, including Bitting midgeCulicoides, LasioheleaLasiohelea, LeptoconopsLeptoconopsWith
Australia's midge categoryAustroconopsDeng.In Bitting midge blood sucking midge, the pathogen propagated by midge matchmaker have blue tongue virus (BTV),
Domestic animal epidemic hemorrhagic fever virus（EHDV）, apply Maron shellfish lattice virus（SBV）, African horse sickness virus（AHSV）Etc. many cause of diseases
Body.Also just because the fields such as blood sucking midge and health care and animal doctor are closely bound up, thus once outburst epizootic will cause it is huge
Economic loss, as Europe if blue tongue disease once occurred（BT）It is sick with Maron shellfish lattice are applied（SB）It is very popular, the country is also in Duo Sheng（Cloud
South, Xinjiang and Inner Mongol etc.）Cattle and sheep in detect BTV.Therefore, precise Identification disease vector is to pass disease to midge to supervise
It surveys, one step of key of prevention.
Currently, difficult in terms of the traditional form of Storehouse midge taxonomic history, one side Storehouse midge ovum, larva and pupa etc. are early
The morphosis of stage of development phase and the polypide of mutilation are difficult to recognize, and Storehouse midge female adult and male worm identify insect degree
There are difference, male to be easy to differentiate using morphological features such as external genital organs and cercoids, and female characteristic of division lacks more difficult differentiation；It is another
Aspect is since the plasticity of Storehouse midge phenotype and the changeability of heredity are also easy to cause erroneous judgement, and traditional form identification operated
Journey is relatively complicated and need to have stronger professional knowledge and abundant practical experience.Therefore, there is an urgent need for seek one kind fast and accurately
Method, to make up the deficiency of traditional form sorting technique.
With the fast development of molecular biology and bioinformatics, using DNA sequence analysis as the Molecular Identification of foundation
As the common technology means of species identification, the defect of traditional taxonomy identification is greatly compensated for.DNA bar code technology refers to
A segment standard, the have variation enough and certain conservative and relatively short DNA fragmentation easily expanded in organism.Therefore, make
For the standard target gene of DNA bar codes, two conditions are must satisfy, first, there must be certain conservative, convenient for logical
With design of primers and carry out large-scale PCR amplification；Second is that have enough variability, to distinguish different plant species, especially closely
Edge kind.
Mitochondria In Developing Flight Muscle of Insects DNA be double-strand closed loop molecule, by ND1-6, ND4L, CO I-CO III, ATPase6, ATPase8,
13 protein genes such as Cytb, 22 tRNA genes and 2 rRNA genes（16S rRNA and 12S rRNA）Totally 37 genes
With the non-coding section composition for including duplication promoter.CO I are the current highest molecular labeling of whole world applying frequency, base
Because size and structure are more conservative, including containing much information, it is easy to be expanded by universal primer again while capable of ensureing to make a variation enough,
There was only the difference of 1%-2% between Different Individual in species, and the difference between sibling species is apparent, and there is more systematic growth
Signal is more suitable for parsing the close biological group of affiliation, is most popular molecular labeling.
So far, having many researchs has shown that I genes of CO are suitable for the Molecular Identification of most biological groups, such as elder brother
The taxonomic identification of the monoids such as worm, birds, fish, nematode.Therefore, using the DNA bar codes techniques of I genes of CO to blood sucking midge into
Row Molecular Identification.This method is proved each other with traditional form sorting technique, can not only realize quick, the accurate mirror of blood sucking midge
Calmly, and using DNA bar code technology it can help to find the cryptic species in Bitting midge or novel species.Currently, having been obtained by this method
It obtains I gene orders of CO of a variety of blood sucking midges and is uploaded in Genbank, but there is no I gene orders of CO of South Mountain Storehouse midge so far.This
The South Mountain Storehouse midge DNA bar code standard gene sequence that invention provides is advantageously implemented quick, the precise Identification of South Mountain Storehouse midge, shortens
The species identification time.
Invention content
Present invention aims at a kind of DNA bar code standard gene sequence of South Mountain Storehouse midge of offer and its Molecular Identification sides
Method.South Mountain Storehouse midge DNA bar code standard gene-I gene orders of CO are obtained by the method for molecular biology DNA sequencing, are passed through
Quick, the precise Identification of South Mountain Storehouse midge are realized in the comparison of gene order similitude.
To achieve the above object, the present invention is achieved through the following technical solutions：
It is prepared by DNA profiling and PCR reacts, by I genes of CO of the primer amplification South Mountain Storehouse midge synthesized, PCR is produced after amplification
Object can detect amplification with agarose gel electrophoresis, and specific amplification is gone out the band that size is about 650bp and send biological public affairs
Department's sequencing.Sequencing result carries out Blast similarity searchings by manual check and correction, sequence assembly in NCBI, it is ensured that gained sequence
It is classified as target gene sequence, while the Identification of Species of South Mountain Storehouse midge relies on Morphological Identification, and is checked through authoritative expert, to protect
The reliability of result is demonstrate,proved.Storehouse midge DNA bar code standard gene in South Mountain of the present invention is I genes of CO, the South Mountain Storehouse midge
I gene orders of CO as shown in SEQ ID No .1：
ctgatccgga ctagttggag ctaccctaag cttattaatt cgaatagaat taagttgccc 60
gggaaatttt tttgctagag accaaattta taatgtaatt gttacttctc atgcatttat 120
tataattttt ttcatagtaa tacctgttat aattggaggc ttcggaaact gattagttcc 180
cttaatacta ggcgccccag acatagcttt cccccgaata aacaatataa gattttgaat 240
acttcccccc tctctttctc ttcttactat tagaagattt gtagacacag gagctggtac 300
tggatgaacg atatacccac ccctctcttc ctttttagcc cacccaaatg cctcagtaga 360
tttagctatt ttttctttac atttagccgg tatctcttcc attttaggag ctgtaaattt 420
tattactaca atcataaata tacgtcctgc aggaataacc ctagataata taccattatt 480
tgtttgatca gtatttatta ctgctattct tttacttctt tctttaccag tactagcagg 540
tgctattacc atattattaa cagatcgaaa tattaatacc tctttttttg accctgccgg 600
aggaggggac cccatccttt atcaacattt attttgattt tttggtcacc 650
The PCR primer of the South Mountain Storehouse midge DNA bar code standard gene sequence, it is characterised in that PCR primer sequence is respectively：
Forward primer：5’TAAACTTCAGGGTGACCAAAAAATCA 3’；
Reverse primer：5’GGTCAACAAATCATAAAGATATTGG 3’.
The method for identifying molecules of the South Mountain Storehouse midge of the present invention, step include：
1）The separation and Extraction DNA from midge tissue to be measured；
2）Using the DNA as template, the primer that uses for：Forward primer：5’TAAACTTCAGGGTGACCAAAAAATCA 3’；Instead
To primer：5 ' GGTCAACAAATCATAAAGATATTGG 3 ' go out I genes of Storehouse midge CO by PCR amplification；
3）Then appropriate step 2 is taken）PCR product detached with agarose gel electrophoresis, sentenced according to electrophoretic band
Whether disconnected is purpose band, if energy specific amplification goes out the band that size is about 650bp, PCR product is cut glue purification, is sent
Biotech firm is sequenced；
4）Analyzed by the comparison of sequencing result, if the similitude of corresponding I gene orders of CO and SEQ ID No .1 98% with
On, you can judge the test serum as South Mountain Storehouse midge.
It is using the advantageous effect of above-mentioned technical proposal：
1）The present invention has combined traditional Morphological Identification and method for identifying molecules, identifies the Storehouse midge, it can be ensured that object
The accuracy and reliability of kind identification.
2）For the present invention compared with traditional Morphological Identification method, the South Mountain Storehouse midge method for identifying molecules established can be significantly
Shorten the qualification time of South Mountain Storehouse midge.
3）The present invention has filled up blank of Storehouse midge CO I gene orders in South Mountain in GenBank databases.
Description of the drawings
Fig. 1 is the techniqueflow chart of the present invention.
Fig. 2 is the electrophoresis pattern of I gene PCR amplifications of South Mountain Storehouse midge CO.Number is described as follows：Swimming lane 1-6 is South Mountain
I genes of CO of Storehouse midge female adult, the band that detection size is about 650bp.M is the DNA marker of Marker III.
Fig. 3 is the electrophoresis pattern of unknown I gene PCR amplifications of Storehouse midge female adult CO.Number is described as follows：Swimming lane 1-2 is
I genes of CO of unknown Storehouse midge female adult, the band that detection size is about 650bp.M is the DNA marker of Marker III.
Specific implementation mode
The acquisition of 1 South Mountain Storehouse midge CO of embodiment, I gene orders
1, the acquisition and preservation of South Mountain Storehouse midge sample
The field habitat that South Mountain Storehouse midge sample sweeps method by net and lamp lures method to be collected in the provinces such as Guizhou, Hainan, Yunnan, and protect
It is stored in 95% alcohol.It is dissected under stereoscope with dissecting needle, Permanent slide is made in remaining in addition to chest, through authoritative special
Family's identification, it is ensured that the accuracy of qualification result.Molecule experiments will be carried out after the independent label of South Mountain Storehouse midge chest.
2, extracting genome DNA
After the South Mountain Storehouse midge chest grinding of preservation, carried out according to QIAGEN DNeasy Blood＆ Tissue Kit specifications
Single Storehouse midge extracting genome DNA, and extremely -20 DEG C of preservation is spare.
3, primer synthesizes
The present embodiment the primer is as follows：
Forward primer：5’TAAACTTCAGGGTGACCAAAAAATCA 3’.
Reverse primer：5’GGTCAACAAATCATAAAGATATTGG 3’.
4, PCR amplification
The PCR reaction systems of the present embodiment are as follows：
DNA profiling 1.5uL
Sense primer（10uM） 1uL
Downstream primer（10uM） 1uL
2×PCR MasterMix 12.5uL
ddH2O 9uL
Total system 25uL
PCR reaction conditions are 94 DEG C of pre-degeneration 5min；94 DEG C of denaturation 1min, 55 DEG C of annealing 30s, 72 DEG C of extension 1min recycle 35
It is secondary；Last 72 DEG C of extensions 5min.
5, PCR product identification and recycling
PCR product determines whether purpose band into row agarose gel electrophoresis, according to electrophoretic band, and electrophoresis pattern is referring to attached drawing
2.PCR product is carried out according to Ago-Gel DNA QIAquick Gel Extraction Kit specifications and cuts glue purification, and product after purification is sent to biological public
Department carries out bidirectional sequencing.
6, CO I genes sequencing
By manually proofread and bioinformatics software South Mountain Storehouse midge CO I gene sequences are compared, edited and spliced after i.e.
For SEQ ID No .1.
The identification of 2 unknown Storehouse midge of embodiment
1, the acquisition and preservation of midge sample
Midge sample sweeps method by net and lamp lures method to be collected in field habitat, and is stored in 95% alcohol.With dissecting needle stereoscopic
It is dissected under mirror, Permanent slide is made for Morphological Identification in remaining in addition to chest.It will be divided after the independent label of midge chest
Son experiment.
2, extracting genome DNA
After the midge chest of preservation is ground, single is carried out not according to QIAGEN DNeasy Blood＆ Tissue Kit specifications
Know midge extracting genome DNA, and extremely -20 DEG C of preservation is spare.
3, primer synthesizes
The present embodiment the primer is as follows：
Forward primer：5’TAAACTTCAGGGTGACCAAAAAATCA 3’.
Reverse primer：5’GGTCAACAAATCATAAAGATATTGG 3’.
4, PCR amplification
The PCR reaction systems of the present embodiment are as follows：
DNA profiling 1.5uL
Sense primer（10uM） 1uL
Downstream primer（10uM） 1uL
2×PCR MasterMix 12.5uL
ddH2O 9uL
Total system 25uL
PCR reaction conditions are 94 DEG C of 5 min of pre-degeneration；94 DEG C of denaturation 1min, 55 DEG C of annealing 30s, 72 DEG C of extension 1min recycle 35
It is secondary；Last 72 DEG C of extensions 5min.
5, PCR product identification and recycling
PCR product determines whether purpose band into row agarose gel electrophoresis, according to electrophoretic band, and electrophoresis pattern is referring to attached drawing
3.Attached drawing 3 shows that the unknown Storehouse midge of embodiment 2 can also go out the product that size is about 650bp by PCR specific amplifications.According to fine jade
Sepharose DNA QIAquick Gel Extraction Kit specifications carry out PCR product and cut glue purification, and product after purification is sent to biotech firm's progress pair
To sequencing.
6, CO I genes sequencing
By manually proofread and bioinformatics software unknown Storehouse midge CO I gene sequences are compared, edited and spliced after with
SEQ ID No .1 carry out similitude comparison, as a result show that the similitude of itself and gene order SEQ ID NO 1 are more than 98%,
It thus can determine that the unknown Storehouse midge is South Mountain Storehouse midge.
Sequence table
<110>Zunyi Medical College
<120>A kind of the DNA bar code standard sequence and its method for identifying molecules of South Mountain Storehouse midge
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 650
<212> DNA
<213>I genes of South Mountain Storehouse midge CO (2 Ambystoma laterale x Ambystoma jeffersonianum)
<400> 1
ctgatccgga ctagttggag ctaccctaag cttattaatt cgaatagaat taagttgccc 60
gggaaatttt tttgctagag accaaattta taatgtaatt gttacttctc atgcatttat 120
tataattttt ttcatagtaa tacctgttat aattggaggc ttcggaaact gattagttcc 180
cttaatacta ggcgccccag acatagcttt cccccgaata aacaatataa gattttgaat 240
acttcccccc tctctttctc ttcttactat tagaagattt gtagacacag gagctggtac 300
tggatgaacg atatacccac ccctctcttc ctttttagcc cacccaaatg cctcagtaga 360
tttagctatt ttttctttac atttagccgg tatctcttcc attttaggag ctgtaaattt 420
tattactaca atcataaata tacgtcctgc aggaataacc ctagataata taccattatt 480
tgtttgatca gtatttatta ctgctattct tttacttctt tctttaccag tactagcagg 540
tgctattacc atattattaa cagatcgaaa tattaatacc tctttttttg accctgccgg 600
aggaggggac cccatccttt atcaacattt attttgattt tttggtcacc 650
<210> 2
<211> 26
<212> DNA
<213>Forward primer (2 Ambystoma laterale x Ambystoma jeffersonianum)
<400> 2
taaacttcag ggtgaccaaa aaatca 26
<210> 3
<211> 25
<212> DNA
<213>Reverse primer (2 Ambystoma laterale x Ambystoma jeffersonianum)
<400> 3
ggtcaacaaa tcataaagat attgg 25
Claims (3)
1. a kind of DNA bar code standard sequence of South Mountain Storehouse midge, it is characterised in that：The DNA bar code standard gene is I bases of CO
Cause is gene order SEQ ID No .1 as described below：
ctgatccgga ctagttggag ctaccctaag cttattaatt cgaatagaat taagttgccc 60
gggaaatttt tttgctagag accaaattta taatgtaatt gttacttctc atgcatttat 120
tataattttt ttcatagtaa tacctgttat aattggaggc ttcggaaact gattagttcc 180
cttaatacta ggcgccccag acatagcttt cccccgaata aacaatataa gattttgaat 240
acttcccccc tctctttctc ttcttactat tagaagattt gtagacacag gagctggtac 300
tggatgaacg atatacccac ccctctcttc ctttttagcc cacccaaatg cctcagtaga 360
tttagctatt ttttctttac atttagccgg tatctcttcc attttaggag ctgtaaattt 420
tattactaca atcataaata tacgtcctgc aggaataacc ctagataata taccattatt 480
tgtttgatca gtatttatta ctgctattct tttacttctt tctttaccag tactagcagg 540
tgctattacc atattattaa cagatcgaaa tattaatacc tctttttttg accctgccgg 600
aggaggggac cccatccttt atcaacattt attttgattt tttggtcacc 650。
2. the PCR primer of the DNA bar code standard gene sequence of South Mountain Storehouse midge described in claim 1, it is characterised in that PCR draws
Object sequence is respectively：
Forward primer：5’TAAACTTCAGGGTGACCAAAAAATCA 3’；
Reverse primer：5’GGTCAACAAATCATAAAGATATTGG 3’.
3. a kind of method for identifying molecules of South Mountain Storehouse midge, step include：
1）The separation and Extraction DNA from midge tissue to be measured；
2）Using the DNA as template, the primer that uses for：Forward primer：5’TAAACTTCAGGGTGACCAAAAAATCA 3’；Instead
To primer：5 ' GGTCAACAAATCATAAAGATATTGG 3 ' go out I genes of Storehouse midge CO by PCR amplification；
3）Then appropriate step 2 is taken）PCR product detached with agarose gel electrophoresis, sentenced according to electrophoretic band
Whether disconnected is purpose band, if energy specific amplification goes out the band that size is about 650bp, PCR product is cut glue purification, is sent
Biotech firm is sequenced；
4）Analyzed by the comparison of sequencing result, if the similitude of corresponding I gene orders of CO and SEQ ID No .1 98% with
On, you can judge the test serum as South Mountain Storehouse midge.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109517909A (en) * | 2019-01-25 | 2019-03-26 | 河北国际旅行卫生保健中心 | A kind of kit and method for identifying Ryukyu's Storehouse midge |
| CN111378764A (en) * | 2020-04-14 | 2020-07-07 | 遵义医科大学 | Identification method of seven species of coe (coe of species) genes based on coe (cytochrome oxidase I) vector culicoides |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102690829A (en) * | 2012-05-16 | 2012-09-26 | 福建国际旅行卫生保健中心 | A DNA bar code standard gene sequence for Taiwan Lasiohelea |
| CN103805609A (en) * | 2014-03-03 | 2014-05-21 | 福建国际旅行卫生保健中心 | Molecular identification method for lasiohelea taiwana |
| CN105483261A (en) * | 2016-01-08 | 2016-04-13 | 中国检验检疫科学研究院 | Specific gene and molecular identification method of culicoides cyancus |
| CN105543249A (en) * | 2016-01-08 | 2016-05-04 | 福建国际旅行卫生保健中心 | Culicoides bellulus specific gene and molecular identification method thereof |
| CN105543364A (en) * | 2016-01-08 | 2016-05-04 | 福建国际旅行卫生保健中心 | Specific gene of Thailand culicoides and molecular identification method thereof |
-
2018
- 2018-01-29 CN CN201810083833.7A patent/CN108486122A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102690829A (en) * | 2012-05-16 | 2012-09-26 | 福建国际旅行卫生保健中心 | A DNA bar code standard gene sequence for Taiwan Lasiohelea |
| CN103805609A (en) * | 2014-03-03 | 2014-05-21 | 福建国际旅行卫生保健中心 | Molecular identification method for lasiohelea taiwana |
| CN105483261A (en) * | 2016-01-08 | 2016-04-13 | 中国检验检疫科学研究院 | Specific gene and molecular identification method of culicoides cyancus |
| CN105543249A (en) * | 2016-01-08 | 2016-05-04 | 福建国际旅行卫生保健中心 | Culicoides bellulus specific gene and molecular identification method thereof |
| CN105543364A (en) * | 2016-01-08 | 2016-05-04 | 福建国际旅行卫生保健中心 | Specific gene of Thailand culicoides and molecular identification method thereof |
Non-Patent Citations (3)
| Title |
|---|
| HARSHA, R等: "KJ909533.2", 《GENBANK》 * |
| 于志超等: "内蒙古阿拉善骆驼生活环境水生双翅目昆虫种类调查", 《昆虫学报》 * |
| 虞以新主编: "《中国蠓科昆虫 第二卷》", 31 March 2006, 军事医学科学出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109517909A (en) * | 2019-01-25 | 2019-03-26 | 河北国际旅行卫生保健中心 | A kind of kit and method for identifying Ryukyu's Storehouse midge |
| CN111378764A (en) * | 2020-04-14 | 2020-07-07 | 遵义医科大学 | Identification method of seven species of coe (coe of species) genes based on coe (cytochrome oxidase I) vector culicoides |
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Application publication date: 20180904 |