CN101492683A - Cultivated silkworm microsporidian polar filament protein PTP3 gene and uses thereof - Google Patents

Abstract

The invention relates to a very important protein gene PTP3 related to propagation and infection of nosema bombycis nageli. A gene order CDS for encoding nosema bombycis nageli polar tube PTP3 is obtained according to 9-fold genome data of the nosema bombycis nageli and proven by a series of biotechnology clonings. The polar tube PTP3 obtained in the invention plays a very important role in studying a propagation and infection mechanism of microsporidian, thus playing a role in productive practice and prevention and treatment of the microsporidian disease. Experiments and analyses such as RNAi intervention and the like indicate that after the polar tube protein PTP3 is intervened, the microsporidian loses the capacity of infecting a host, therefore, the polar tube protein PTP3 of the microsporidian and potential regulator genes thereof become new factors for treating microsporidian infection, in addition, the method of modifying genes can be adopted to express the polar tube protein PTP3 in an abundant way, thus killing part of pests such as locust and the like.

Description

Cultivated silkworm microsporidian polar filament protein PTP 3 gene and purposes

Technical field

This discovery relates to biological technical field, particularly relates to a kind of breeding that comes from nosema bombycis and infects relevant important protein PTP 3 gene.

Background technology

Microsporidium is widespread distribution obligatory parasitism protozoon in natural cell, kind surplus nearly 150 genus 1200, host very widely being arranged, comprise vertebrates and invertebrates, is the fatal cause of disease of economic insects, fish, Lagomorpha, product hair animal, rodents and primates.And nosema bombycis (Nosema bombycis), classification position belongs to Protozoa, monofilament Sporozoa, Microsporidea.Parasitize the economic insects silkworm and cause that serious prion is sick by microsporidium, and, classified as silkworm by the sericulture countries and regions always and already produce legal Quarantine Objects because little spore protozoon can be propagated by the embryo in downward generation.List promptly reaches more than the up to ten million units because of the direct economic loss of being with seed culture of viruses and pebrine to bring with regard to China every year.Capturing the chronic disease of this silk industry of pebrine, is the emphasis and the difficult point research field of silkworm industry.

Pebrine is the disease that is caused by the microsporidium parasitic host.Microsporidium is with its unique mode of infection, extensively parasitic and infection has insect, fish, rodents, Lagomorpha, fleece animal, primates and household pet such as the dog and the bird of economic worth, cause serious disease even death, seriously restricting world silkworm and already producing and the animal child care.Even can also opportunistic ground aids infection (HIV) patient, organ transplantation patient, take immune suppressant drug patient and innate immunity incomcopetence person, cause malignant diseases such as respiratory system, urinary system and skin ulceration, aggravate one's illness, quicken patient's death.Over more than 100 year, although used the number of chemical medicine to treat, as: imidazoles, fumidil, song miaow pyridine alkali, benzene Lai Te, thiophanate methyl or the like, result of treatment is unsatisfactory, finds to have the specific medicament of treatment microsporidiosis so far not yet.

Utmost point pipe is nosema bombycis breeding and the vitals that infect, plays key effect nosema bombycis in the infecting of silkworm.Utmost point length of tube is generally at 50～150 μ m, and diameter is generally at 0.1～0.2 mu m range.Utmost point pipe has snappiness, and its pipe diameter can be increased to 0.4 μ m in transporting sporoplasmic process, and sporoplasm transports its length of back that finishes can shorten 5～10%.In the nosema bombycis phagocytic process, utmost point pipe injects sporoplasm its tenuigenin simultaneously suitably stimulating rapid ejection the in back also " to thrust " in the bombyx mori cell, finishes the invasion to silkworm.And its moiety mainly is an albumen, and the polar tube protein (ptp) PTP1 main integral part that is it.Polar tube protein (ptp) PTP1 is a kind of concanavalin A, is the major protein of utmost point pipe, and the proteic content of PTP1 accounts for 70% of whole polar tube protein (ptp) in E.hellom.Dye analysis PTP1 by SDA-PAGE and silver and find that the PTP1 molecular size of different types of microsporidium does not wait.To proteic studies show that of different microsporidium PTP1, PTP1 albumen has hydrophobicity, high proline content and immunizing antigen etc. such as feature (Delbac et al., 1998).And PTP2 and PTP3 also are the important component that constitutes utmost point pipe.Have research to think, in utmost point pipe ejection process, PTP1 and PTP2 were class crystalloid protein monomers before spore activates, and form polymer in the germination process on utmost point pipe top, and PTP3 influenced the ejection of utmost point pipe by the polymeric conformational state of regulation and control PTP1-PTP2.In addition, the glycosylation of polar tube protein (ptp) PTP1 plays an important role to the structure of utmost point pipe, thereby the remote effect microsporidium is to the invasion of host cell.But the research about the microsporidium polar tube protein (ptp) only is confined to a small amount of microsporidium that part microsporidiums such as Encephalitozoon, Glugea, Ameson and Nosema belong at present.

Summary of the invention

The present invention intends based on the genome analysis data, and its purpose is to provide a kind of nosema bombycis polar tube protein (ptp) PTP3 gene.By analyzing function, composition and the characteristic of PTP3, understand fully the effect of polar tube protein (ptp) PTP3 in nosema bombycis utmost point pipe ejection process, verify nosema bombycis and infect the morbific mechanism of host silkworm, production practice and diseases prevention and treatment are extremely important.

The present patent application people has obtained the encoding sequence of this protein gene by the clone to cultivated silkworm microsporidian polar filament protein PTP 3 gene, and concrete grammar is as follows:

(1) homology prediction:, obtained nosema bombycis polar tube protein (ptp) PTP3 gene order CDS by to nosema bombycis and the genomic dna homolog analysis of rabbit encephalitis microsporidium;

(2) design of primers: utilize primer-design software Primer 5.0 to carry out design of primers, add the BamHI restriction enzyme site at forward primer, add Sal I restriction enzyme site at reverse primer, primer sequence is respectively:

PTP3-Primer-F5’-CGGGATCCCTACACATTATTTG-3’

PTP3-Primer-R5’-CGGTCGACGTGATGATGGGATTCC-3’

(3) polar tube protein PTP3 pcr amplification and clone: utilize the little spore genome that extracts directly as template, adopt genome pcr amplification purpose fragment; Amplified fragments is cloned on pMD18-T (TaKaRa) carrier, carries out sequencing (Shanghai is handsome) then, thus the polar tube protein PTP3 complete encoding sequence that obtains;

(4) sequence homology analysis: the homology comparison is finished with BLAST on the NCBI website, protein does not have significant homology in BLAST analysis revealed cultivated silkworm microsporidian polar filament protein PTP 3 and the NCBI protein library, and polar tube protein PTP3 is the important albumen of nosema bombycis utmost point pipe, so polar tube protein PTP3 gene is a kind of newfound protein gene.

The PTP3 full length gene cDNA gene that aforesaid method obtains and the aminoacid sequence of deduction thereof are as follows:

atgatctacacattatttgtaatatacatatctagcacattttgt

M I Y T L F V I Y I S S T F C

aggataatctcaagaagtacagtagaaaatggtcctcccacagtt

R I I S R S T V E N G P P T V

cctttcgatgaacatcaagctaaacatgttcatgttcgagtgcac

P F D E H Q A K H V H V R V H

ggtagacacagacatggaaatcaccatggtggtggtcaagacgta

G R H R H G N H H G G G Q D V

cacgaagttcaaaaacaccacagtcatcaccacaatgggcataat

H E V Q K H H S H H H N G H N

tacggtcttgaacatggaaaattcggaaatgtaacttacacttct

Y G L E H G K F G N V T Y T S

ggaaatcaccatggtcaccattctcatcacggttctcacgcggat

G N H H G H H S H H G S H A D

ggacaccattctcaatacggttcgcacgcggacggacatcatgtt

G H H S Q Y G S H A D G H H V

aacgcccaatcagatgcttctgcaccactagacgttttgccaatg

N A Q S D A S A P L D V L P M

aaagaagtaagaagtggtgatgatccagctttggagagagatcta

K E V R S G D D P A L E R D L

ggaattcaacaagccatgaaagcttttcgatcttctttacctgtt

G I Q Q A M K A F R S S L P V

tcaaataaagacaccaattctcttttggataaagctgaaggaatt

S N K D T N S L L D K A E G I

ttaagaagtgcaccagacggccaattagatgctcctgttcttgat

L R S A P D G Q L D A P V L D

agttacggaaatccaactgggcagacatacagagaagccgcagaa

S Y G N P T G Q T Y R E A A E

gatgcagtaggctataagtatccaacagcagtagcagatgaagaa

D A V G Y K Y P T A V A D E E

ttcgatagtttagtaaggagtggtgttgaaccgggaagagccaag

F D S L V R S G V E P G R A K

caagtagctgataaaattgaagaaacaggttttgaacacgagaaa

Q V A D K I E E T G F E H E K

attaaagatgcagtaagaaatgaagtagctagaggtacagcaaac

I K D A V R N E V A R G T A N

agagccgtagcagaattacaagacgaaattcaaagacgtaaaaat

R A V A E L Q D E I Q R R K N

gctactaaagtcaaagaacgtgttaaagttgaagatgaattgtac

A T K V K E R V K V E D E L Y

agacaacacactcttcctgctactggtttattcgatgaagacgtg

R Q H T L P A T G L F D E D V

tctgttctgaaaaatatgagccaaattgttccgcctgcgaaaatt

S V L K N M S Q I V P P A K I

gtcgactacatcagagactcctatgataattcaattgaagccgga

V D Y I R D S Y D N S I E A G

cttcctcctgcggttgctcgtgctcaggccgatcgtgttggttct

L P P A V A R A Q A D R V G S

ttaattacggatttcaatacaatgttaagaagtggtgaatctcct

L I T D F N T M L R S G E S P

gaaactgctttgtcccatcttaaagacatgtctgataggcaatca

E T A L S H L K D M S D R Q S

aaagtcgatcaaagaagagaattacttcagaactctagtccagaa

K V D Q R R E L L Q N S S P E

gtcttaaaaatactttcagataaagataatttaattaaagaagca

V L K I L S D K D N L I K E A

ctggttaaaactgcaagagatcctgactgtagagacagatacttt

L V K T A R D P D C R D R Y F

gatgaagcctcccagaagatgaagattcttgatgcaagagaagcc

D E A S Q K M K I L D A R E A

agttttattcatgatactgaaaataacaagcacatacgtcgtcct

S F I H D T E N N K H I R R P

aggaaaatggcagaagatttaatggataagactaacagagataat

R K M A E D L M D K T N R D N

gcagaattagttgatgtagcttatagtcacccagtagtagccaaa

A E L V D V A Y S H P V V A K

gacattttaaaacaaggtattgcaaacactgaaagagtagctgag

D I L K Q G I A N T E R V A E

aaattggctgatcatttggatgctgtaaaatttgctgaaaaagct

K L A D H L D A V K F A E K A

caaaatccttcagctgtttcggctttgaaacttcttgacgagaga

Q N P S A V S A L K L L D E R

aagagacaagatgaacttgaaaaaatcaataagagaaatacagaa

K R Q D E L E K I N K R N T E

aatgagcaaaagatgttcaagcagattctaaaagaccaagaagac

N E Q K M F K Q I L K D Q E D

gcggaagtaatgcaaaccgaagagaaaatggcagccgcaaacaaa

A E V M Q T E E K M A A A N K

gcaagaaagattttagatgaaggacgtagggcagctgaacaagtt

A R K I L D E G R R A A E Q V

ttgaaagcgggtggaagtactgatgaagctgctgctgttcgtgtt

L K A G G S T D E A A A V R V

gcagctgagaatgctactgctcgaggtttggaacaaagagaatcc

A A E N A T A R G L E Q R E S

atgatcgaagaagagattatgaaaaggtttggaggcgcaggtgat

M I E E E I M K R F G G A G D

cctgaaaaagctcgtaaaatggctgctcacattgctatcgctcac

P E K A R K M A A H I A I A H

agtggagaaaatgccgtaggaaaaacgcccgaagaagtcattaag

S G E N A V G K T P E E V I K

aaagaacaagaagcgcaaaagaaagagattttaagaaatgctttg

K E Q E A Q K K E I L R N A L

tatgatgatattggaaaagaaccaatcgaaattaagtctgtaatt

Y D D I G K E P I E I K S V I

ccagaagaccgagatgaatactctattgacagaagtgtaattaaa

P E D R D E Y S I D R S V I K

tttcctcttaaaacatttattgatgaagaagtaagcaatgttgga

F P L K T F I D E E V S N V G

gaagcctacgtaaagaacaaaaagcaaagtatcaacgacgaagtc

E A Y V K N K K Q S I N D E V

aggaataaagtcactaccaccaacgtcaacagcggtcttaatgta

R N K V T T T N V N S G L N V

attgaaacagaaaatggtttcttgaatctaggagagaatagtaag

I E T E N G F L N L G E N S K

aaaatccacattgacgaagccacagcctattttaaaccggccgcc

K I H I D E A T A Y F K P A A

aaattaaaaatggagaaaaaaggtattaagacggataatttcaga

K L K M E K K G I K T D N F R

ccaaaaaccaatcaaggtgaaataagagacatgcccaaaggagaa

P K T N Q G E I R D M P K G E

acttactatgaggttgatcttaaagacgccgatctaagtcttcct

T Y Y E V D L K D A D L S L P

tctcctactaatcctactcctgatacacttgtctcaaatggaaaa

S P T N P T P D T L V S N G K

gacgttgttgatgttgaagatgtttctgccattgtaattaataaa

D V V D V E D V S A I V I N K

ggtactttagtccagacaccatggaacgaatactctgacatgatt

G T L V Q T P W N E Y S D M I

aagccgaagtttaatccgtatccaaatgaaggagagaaaattaat

K P K F N P Y P N E G E K I N

caaatcaagaaacttcaaaaactcatagccgatgagaagagaaaa

Q I K K L Q K L I A D E K R K

gacacattagaccgaattaaagttaacacaattactaatcctgac

D T L D R I K V N T I T N P D

ggagaaaagcgaatgattgttaatacgccttatggtgttcaagaa

G E K R M I V N T P Y G V Q E

atgttcgaggaaactgaaggaatgaaaagactttctcatattgat

M F E E T E G M K R L S H I D

ccaaataagaacgtggctattagtgaaactccgactgatgataac

P N K N V A I S E T P T D D N

aaagaatctatttacagcgcctttaagaacgtttcaccaaatgaa

K E S I Y S A F K N V S P N E

gcagtcggaaaattcattgaagatacttttaattctgcatactca

A V G K F I E D T F N S A Y S

tccggtcaagctcagagatttgtaaatcccacaaacgcctttact

S G Q A Q R F V N P T N A F T

ggtcaagaagatccaaagaaagccagactcatgactcagaagact

G Q E D P K K A R L M T Q K T

attgatgttacagagtattatattaatgacggtaaaccatcttcg

I D V T E Y Y I N D G K P S S

gctatcaaaaatcaactaatccaaaattacagagccttagctgat

A I K N Q L I Q N Y R A L A D

agccttggaatgaccgaagaagatttcattcaattcgctcgtaaa

S L G M T E E D F I Q F A R K

attcctgatgattctttggcacaaatgatctcttataatgaacaa

I P D D S L A Q M I S Y N E Q

tcagaatcttcgaggcccgcattagtcgatgctccattctttaag

S E S S R P A L V D A P F F K

acacttcagccacttgcaaatcaaacgtcaaagaccaaacttaac

T L Q P L A N Q T S K T K L N

gacatcatcaaaataatcttcacacaaatttctaaagtaactggt

D I I K I I F T Q I S K V T G

aaaaccaattcaaccacaggcacaacactcgagaagaagatagtt

K T N S T T G T T L E K K I V

cctaatctaaaggcagtaagatctgaccaagtcattgcaggacca

P N L K A V R S D Q V I A G P

aatggaggaacttcagctttatcccaagccacagctccaagaacc

N G G T S A L S Q A T A P R T

ggttctacagttttgagtaagcaaatcacacgaggacttcccaga

G S T V L S K Q I T R G L P R

gttccttcaggtacaggtactagttatccagtgagagatcccaat

V P S G T G T S Y P V R D P N

ggtattaatacaagtgaagataatgaaagatataaagtcaatcct

G I N T S E D N E R Y K V N P

tataatccatattctaaatctgatactagtggtcttgaagcacct

Y N P Y S K S D T S G L E A P

ggtggtgaaatagtacataatggtacaagaccaagtccttacgct

G G E I V H N G T R P S P Y A

atagtaggagttccaatagtcgcagcggttacaacaccagtcatc

I V G V P I V A A V T T P V I

agaccagcagtcttaagaacgcctccagcagcacaatcgagttct

R P A V L R T P P A A Q S S S

agtgcacttcaaggaaatacagcacttactggagcaggaacacca

S A L Q G N T A L T G A G T P

agagccggagtagcaggatcaccaggagtaaatccaggaccaact

R A G V A G S P G V N P G P T

ggtaaagcacctgtctcaccccctgcaccaacatcaccagcttca

G K A P V S P P A P T S P A S

tctcctacagcctcacctatagcacctacagcttcacctatagca

S P T A S P I A P T A S P I A

cctacaggagcatcaataggagcacctggttcacttggaatccca

P T G A S I G A P G S L G I P

tcatcaccattgaag

S S P L K

The PTP3 gene particularly kills application in the locust agent at the preparation sterilant.

Advantage of the present invention is:

(1) because microsporidium polar tube protein (ptp) PTP3 is a major protein of forming microsporidium utmost point pipe, and utmost point pipe is the vitals that microsporidium is bred and infected, insect, fish, rodents, Lagomorpha, fleece animal, primates and the household pet such as dog and the bird that have economic worth by its extensively parasitic also infection, cause serious disease even death, seriously restricting world silkworm and already producing and the animal child care.Even can also opportunistic ground aids infection (HIV) patient, organ transplantation patient, take immune suppressant drug patient and innate immunity incomcopetence person, cause malignant diseases such as respiratory system, urinary system and skin ulceration, aggravate one's illness, quicken patient's death.Therefore the polar tube protein (ptp) PTP3 that is obtained has its vital role to breeding of research microsporidium and infection mechanism.By analyzing function, composition and the characteristic of PTP3, understand fully the effect of polar tube protein (ptp) PTP3 in nosema bombycis utmost point pipe ejection process, verify nosema bombycis and infect the morbific mechanism of host silkworm, production practice and diseases prevention and treatment are extremely important.

(2) analysis of experiments such as RNAi interference shows that after polar tube protein PTP3 was interfered with, microsporidium lost the ability of infection host.So will becoming the treatment microsporidium, the polar tube protein PTP3 of microsporidium and its potential regulatory gene infect new factor.Can also make it its abundant expression by genetically modified means, thereby murder part insects such as locust.

Embodiment:

Use Glimmer3.0 software,, obtained nosema bombycis polar tube protein (ptp) PTP3 gene order CDS by to nosema bombycis and the genomic dna homolog analysis of rabbit encephalitis microsporidium; Use Primer5.0 software, design a pair of Auele Specific Primer

PTP3-Primer-F5’-CGGGATCCCTACACATTATTTG-3’

PTP3-Primer-R5’-CGGTCGACGTGATGATGGGATTCC-3’

The amplification segment is cloned on the pMD18-T carrier, and sequencing is finished by Shanghai English fine horse, the polar tube protein PTP3 complete encoding sequence of acquisition; The homology comparison is finished with BLAST on the NCBI website, and sequential analysis shows that it is a new protein gene.

Sequence table

SEQUENCE?LISTING

＜110〉Southwestern University

＜120〉cultivated silkworm microsporidian polar filament protein PTP 3 gene

<130>

<140>

<141>

<160>1

<170>

<210>1

<211>4110

<212>DNA

＜213〉silkworm (Bombyx mori)

<400>1

1 atgatctaca?cattatttgt?aatatacata?tctagcacat?tttgtaggat?aatctcaaga

61 agtacagtag?aaaatggtcc?tcccacagtt?cctttcgatg?aacatcaagc?taaacatgtt

121 catgttcgag?tgcacggtag?acacagacat?ggaaatcacc?atggtggtgg?tcaagacgta

181 cacgaagttc?aaaaacacca?cagtcatcac?cacaatgggc?ataattacgg?tcttgaacat

241 ggaaaattcg?gaaatgtaac?ttacacttct?ggaaatcacc?atggtcacca?ttctcatcac

301 ggttctcacg?cggatggaca?ccattctcaa?tacggttcgc?acgcggacgg?acatcatgtt

361 aacgcccaat?cagatgcttc?tgcaccacta?gacgttttgc?caatgaaaga?agtaagaagt

421 ggtgatgatc?cagctttgga?gagagatcta?ggaattcaac?aagccatgaa?agcttttcga

481 tcttctttac?ctgtttcaaa?taaagacacc?aattctcttt?tggataaagc?tgaaggaatt

541 ttaagaagtg?caccagacgg?ccaattagat?gctcctgttc?ttgatagtta?cggaaatcca

601 actgggcaga?catacagaga?agccgcagaa?gatgcagtag?gctataagta?tccaacagca

661 gtagcagatg?aagaattcga?tagtttagta?aggagtggtg?ttgaaccggg?aagagccaag

721 caagtagctg?ataaaattga?agaaacaggt?tttgaacacg?agaaaattaa?agatgcagta

781 agaaatgaag?tagctagagg?tacagcaaac?agagccgtag?cagaattaca?agacgaaatt

841 caaagacgta?aaaatgctac?taaagtcaaa?gaacgtgtta?aagttgaaga?tgaattgtac

901 agacaacaca?ctcttcctgc?tactggttta?ttcgatgaag?acgtgtctgt?tctgaaaaat

961 atgagccaaa?ttgttccgcc?tgcgaaaatt?gtcgactaca?tcagagactc?ctatgataat

1021?tcaattgaag?ccggacttcc?tcctgcggtt?gctcgtgctc?aggccgatcg?tgttggttct

1081?ttaattacgg?atttcaatac?aatgttaaga?agtggtgaat?ctcctgaaac?tgctttgtcc

1141?catcttaaag?acatgtctga?taggcaatca?aaagtcgatc?aaagaagaga?attacttcag

1201?aactctagtc?cagaagtctt?aaaaatactt?tcagataaag?ataatttaat?taaagaagca

1261?ctggttaaaa?ctgcaagaga?tcctgactgt?agagacagat?actttgatga?agcctcccag

1321?aagatgaaga?ttcttgatgc?aagagaagcc?agttttattc?atgatactga?aaataacaag

1381?cacatacgtc?gtcctaggaa?aatggcagaa?gatttaatgg?ataagactaa?cagagataat

1441?gcagaattag?ttgatgtagc?ttatagtcac?ccagtagtag?ccaaagacat?tttaaaacaa

1501?ggtattgcaa?acactgaaag?agtagctgag?aaattggctg?atcatttgga?tgctgtaaaa

1561?tttgctgaaa?aagctcaaaa?tccttcagct?gtttcggctt?tgaaacttct?tgacgagaga

1621?aagagacaag?atgaacttga?aaaaatcaat?aagagaaata?cagaaaatga?gcaaaagatg

1681?ttcaagcaga?ttctaaaaga?ccaagaagac?gcggaatgaa?tgcaaaccga?agagaaaatg

1741?gcagccgcaa?acaaagcaag?aaagatttta?gatgaaggac?gtagggcagc?tgaacaagtt

1801?ttgaaagcgg?gtggaagtac?tgatgaagct?gctgctgttc?gtgttgcagc?tgagaatgct

1861?actgctcgag?gtttggaaca?aagagaatcc?atgatcgaag?aagagattat?gaaaaggttt

1921?ggaggcgcag?gtgatcctga?aaaagctcgt?aaaatggctg?ctcacattgc?tatcgctcac

1981?agtggagaaa?atgccgtagg?aaaaacgccc?gaagaagtca?ttaagaaaga?acaagaagcg

2041?caaaagaaag?agattttaag?aaatgctttg?tatgatgata?ttggaaaaga?accaatcgaa

2101?attaagtctg?taattccaga?agaccgagat?gaatactcta?ttgacagaag?tgtaattaaa

2161?tttcctctta?aaacatttat?tgatgaagaa?gtaagcaatg?ttggagaagc?ctacgtaaag

2221?aacaaaaagc?aaagtatcaa?cgacgaagtc?aggaataaag?tcactaccac?caacgtcaac

2281?agcggtctta?atgtaattga?aacagaaaat?ggtttcttga?atctaggaga?gaatagtaag

2341?aaaatccaca?ttgacgaagc?cacagcctat?tttaaaccgg?ccgccaaatt?aaaaatggag

2401?aaaaaaggta?ttaagacgga?taatttcaga?ccaaaaacca?atcaaggtga?aataagagac

2461?atgcccaaag?gagaaactta?ctatgaggtt?gatcttaaag?acgccgatct?aagtcttcct

2521?tctcctacta?atcctactcc?tgatacactt?gtctcaaatg?gaaaagacgt?tgttgatgtt

2581?gaagatgttt?ctgccattgt?aattaataaa?ggtactttag?tccagacacc?atggaacgaa

2641?tactctgaca?tgattaagcc?gaagtttaat?ccgtatccaa?atgaaggaga?gaaaattaat

2701?caaatcaaga?aacttcaaaa?actcatagcc?gatgagaaga?gaaaagacac?attagaccga

2761?attaaagtta?acacaattac?taatcctgac?ggagaaaagc?gaatgattgt?taatacgcct

2821?tatggtgttc?aagaaatgtt?cgaggaaact?gaaggaatga?aaagactttc?tcatattgat

2881?ccaaataaga?acgtggctat?tagtgaaact?ccgactgatg?ataacaaaga?atctatttac

2941?agcgccttta?agaacgtttc?accaaatgaa?gcagtcggaa?aattcattga?agatactttt

3001?aattctgcat?actcatccgg?tcaagctcag?agatttgtaa?atcccacaaa?cgcctttact

3061?ggtcaagaag?atccaaagaa?agccagactc?atgactcaga?agactattga?tgttacagag

3121?tattatatta?atgacggtaa?accatcttcg?gctatcaaaa?atcaactaat?ccaaaattac

3181?agagccttag?ctgatagcct?tggaatgacc?gaagaagatt?tcattcaatt?cgctcgtaaa

3241?attcctgatg?attctttggc?acaaatgatc?tcttataatg?aacaatcaga?atcttcgagg

3301?cccgcattag?tcgatgctcc?attctttaag?acacttcagc?cacttgcaaa?tcaaacgtca

3361?aagaccaaac?ttaacgacat?catcaaaata?atcttcacac?aaatttctaa?agtaactggt

3421?aaaaccaatt?caaccacagg?cacaacactc?gagaagaaga?tagttcctaa?tctaaaggca

3481?gtaagatctg?accaagtcat?tgcaggacca?aatggaggaa?cttcagcttt?atcccaagcc

3541?acagctccaa?gaaccggttc?tacagttttg?agtaagcaaa?tcacacgagg?acttcccaga

3601?gttccttcag?gtacaggtac?tagttatcca?gtgagagatc?ccaatggtat?taatacaagt

3661?gaagataatg?aaagatata agtcaatcct?tataatccat?attctaaatc?tgatactagt

3721?ggtcttgaag?cacctggtgg?tgaaatagta?cataatggta?caagaccaag?tccttacgct

3781?atagtaggag?ttccaatagt?cgcagcggtt?acaacaccag?tcatcagacc?agcagtctta

3841?agaacgcctc?cagcagcaca?atcgagttct?agtgcacttc?aaggaaatac?agcacttact

3901?ggagcaggaa?caccaagagc?cggagtagca?ggatcaccag?gagtaaatcc?aggaccaact

3961?ggtaaagcac?ctgtctcacc?ccctgcacca?acatcaccag?cttcatctcc?tacagcctca

4021?cctatagcac?ctacagcttc?acctatagca?cctacaggag?catcaatagg?agcacctggt

4081?tcacttggaa?tcccatcatc?accattgaag

<210>2

<211>1370

<212>PRT

＜213〉silkworm (Bombyx mori)

<400>1

1 MIYTLFVIYI?SSTFCRIISR?STVENGPPTV?PFDEHQAKHV?HVRVHGRHRH

GNHHGGGQDV

61 HEVQKHHSHH?HNGHNYGLEH?GKFGNVTYTS?GNHHGHHSHH

GSHADGHHSQ?YGSHADGHHV

121?NAQSDASAPL?DVLPMKEVRS?GDDPALERDL?GIQQAMKAFR?SSLPVSNKDT

NSLLDKAEGI

181?LRSAPDGQLD?APVLDSYGNP?TGQTYREAAE?DAVGYKYPTA?VADEEFDSLV

RSGVEPGRAK

241?QVADKIEETG?FEHEKIKDAV?RNEVARGTAN?RAVAELQDEI?QRRKNATKVK

ERVKVEDELY

301 RQHTLPATGL?FDEDVSVLKN?MSQIVPPAKI?VDYIRDSYDN?SIEAGLPPAV

ARAQADRVGS

361 LITDFNTMLR?SGESPETALS?HLKDMSDRQS?KVDQRRELLQ?NSSPEVLKIL

SDKDNLIKEA

421 LVKTARDPDC?RDRYFDEASQ?KMkiLDAREA?SFIHDTENNK?HIRPRKMAE

DLMDKTNRDN

481 AELVDVAYSH?PVVAKDILKQ?GIANTERVAE?KLADHLDAVK?FAEKAQNPSA

VSALKLLDER

541 KRQDELEKIN?KRNTENEQKM?FKQILKDQED?AEVMQTEEKM?AAANKARKIL

DEGRRAAEQV

601 LKAGGSTDEA?AAVRVAAENA?TARGLEQRES?MIEEEIMKRF?GGAGDPEKAR

KMAAHIAIAH

661 SGENAVGKTP?EEVIKKEQEA?QKKEILRNAL?YDDIGKEPIE?IKSVIPEDRD

EYSIDRSVIK

721 FPLKTFIDEE?VSNVGEAYVK?NKKQSINDEV?RNKVTTTNVN?SGLNVIETEN

GFLNLGENSK

781 KIHIDEATAY?FKPAAKLKME?KKGIKTDNFR?PKTNQGEIRD?MPKGETYYEV

DLKDADLSLP

841 SPTNPTPDTL?VSNGKDVVDV?EDVSAIVINK?GTLVQTPWNE?YSDMIKPKFN

PYPNEGEKIN

901 QIKKLQKLIA?DEKRKDTLDR?IKVNTITNPD?GEKRMIVNTP?YGVQEMFEET

EGMKRLSHID

961 PNKNVAISET?PTDDNKESIY?SAFKNVSPNE?AVGKFIEDTF?NSAYSSGQAQ

RFVNPTNAFT

1021?GQEDPKKARL?MTQKTIDVTE?YYINDGKPSS?AIKNQLIQNY?RALADSLGMT

EEDFIQFARK

1081?IPDDSLAQMI?SYNEQSESSR?PALVDAPFFK?TLQPLANQTS?KTKLNDIIKI

IFTQISKVTG

1141?KTNSTTGTTL?EKKIVPNLKA?VRSDQVIAGP?NGGTSALSQA?TAPRTGSTVL

SKQITRGLPR

1201?VPSGTGTSYP?VRDPNGINTS?EDNERYKVNP?YNPYSKSDTS?GLEAPGGEIV

HNGTRPSPYA

1261?IVGVPIVAAV?TTPVIRPAVL?RTPPAAQSSS?SALQGNTALT?GAGTPRAGVA

GSPGVNPGPT

1321?GKAPVSPPAP?TSPASSPTAS?PIAPTASPIA?PTGASIGAPG?SLGIPSSPLK

Claims (3)

1, a kind of cultivated silkworm microsporidian polar filament protein PTP 3 gene is characterized in that its sequence is as follows:

1?atgatctaca?cattatttgt?aatatacata?tctagcacat?tttgtaggat?aatctcaaga

61?agtacagtag?aaaatggtcc?tcccacagtt?cctttcgatg?aacatcaagc?taaacatgtt

121?catgttcgag?tgcacggtag?acacagacat?ggaaatcacc?atggtggtgg?tcaagacgta

181?cacgaagttc?aaaaacacca?cagtcatcac?cacaatgggc?ataattacgg?tcttgaacat

241?ggaaaattcg?gaaatgtaac?ttacacttct?ggaaatcacc?atggtcacca?ttctcatcac

301?ggttctcacg?cggatggaca?ccattctcaa?tacggttcgc?acgcggacgg?acatcatgtt

361?aacgcccaat?cagatgcttc?tgcaccacta?gacgttttgc?caatgaaaga?agtaagaagt

421?ggtgatgatc?cagctttgga?gagagatcta?ggaattcaac?aagccatgaa?agcttttcga

481?tcttctttac?ctgtttcaaa?taaagacacc?aattctcttt?tggataaagc?tgaaggaatt

541?ttaagaagtg?caccagacgg?ccaattagat?gctcctgttc?ttgatagtta?cggaaatcca

601?actgggcaga?catacagaga?agccgcagaa?gatgcagtag?gctataagta?tccaacagca

661?gtagcagatg?aagaattcga?tagtttagta?aggagtggtg?ttgaaccggg?aagagccaag

721?caagtagctg?ataaaattga?agaaacaggt?tttgaacacg?agaaaattaa?agatgcagta

781?agaaatgaag?tagctagagg?tacagcaaac?agagccgtag?cagaattaca?agacgaaatt

841?caaagacgta?aaaatgctac?taaagtcaaa?gaacgtgtta?aagttgaaga?tgaattgtac

901?agacaacaca?ctcttcctgc?tactggttta?ttcgatgaag?acgtgtctgt?tctgaaaaat

961?atgagccaaa?ttgttccgcc?tgcgaaaatt?gtcgactaca?tcagagactc?ctatgataat

1021?tcaattgaag?ccggacttcc?tcctgcggtt?gctcgtgctc?aggccgatcg?tgttggttct

1081?ttaattacgg?atttcaatac?aatgttaaga?agtggtgaat?ctcctgaaac?tgctttgtcc

1141?catcttaaag?acatgtctga?taggcaatca?aaagtcgatc?aaagaagaga?attacttcag

1201?aactctagtc?cagaagtctt?aaaaatactt?tcagataaag?ataatttaat?taaagaagca

1261?ctggttaaaa?ctgcaagaga?tcctgactgt?agagacagat?actttgatga?agcctcccag

1321?aagatgaaga?ttcttgatgc?aagagaagcc?agttttattc?atgatactga?aaataacaag

1381?cacatacgtc?gtcctaggaa?aatggcagaa?gatttaatgg?ataagactaa?cagagataat

1441?gcagaattag?ttgatgtagc?ttatagtcac?ccagtagtag?ccaaagacat?tttaaaacaa

1501?ggtattgcaa?acactgaaag?agtagctgag?aaattggctg?atcatttgga?tgctgtaaaa

1561?tttgctgaaa?aagctcaaaa?tccttcagct?gtttcggctt?tgaaacttct?tgacgagaga

1621?aagagacaag?atgaacttga?aaaaatcaat?aagagaaata?cagaaaatga?gcaaaagatg

1681?ttcaagcaga?ttctaaaaga?ccaagaagac?gcggaagtaa?tgcaaaccga?agagaaaatg

1741?gcagccgcaa?acaaagcaag?aaagatttta?gatgaaggac?gtagggcagc?tgaacaagtt

1801?ttgaaagcgg?gtggaagtac?tgatgaagct?gctgctgttc?gtgttgcagc?tgagaatgct

1861?actgctcgag?gtttggaaca?aagagaatcc?atgatcgaag?aagagattat?gaaaaggttt

1921?ggaggcgcag?gtgatcctga?aaaagctcgt?aaaatggctg?ctcacattgc?tatcgctcac

1981?agtggagaaa?atgccgtagg?aaaaacgccc?gaagaagtca?ttaagaaaga?acaagaagcg

2041?caaaagaaag?agattttaag?aaatgctttg?tatgatgata?ttggaaaaga?accaatcgaa

2101?attaagtctg?taattccaga?agaccgagat?gaatactcta?ttgacagaag?tgtaattaaa

2161?tttcctctta?aaacatttat?tgatgaagaa?gtaagcaatg?ttggagaagc?ctacgtaaag

2221?aacaaaaagc?aaagtatcaa?cgacgaagtc?aggaataaag?tcactaccac?caacgtcaac

2281?agcggtctta?atgtaattga?aacagaaaat?ggtttcttga?atctaggaga?gaatagtaag

2341?aaaatccaca?ttgacgaagc?cacagcctat?tttaaaccgg?ccgccaaatt?aaaaatggag

2401?aaaaaaggta?ttaagacgga?taatttcaga?ccaaaaacca?atcaaggtga?aataagagac

2461?atgcccaaag?gagaaactta?ctatgaggtt?gatcttaaag?acgccgatct?aagtcttcct

2521?tctcctacta?atcctactcc?tgatacactt?gtctcaaatg?gaaaagacgt?tgttgatgtt

2581?gaagatgttt?ctgccattgt?aattaataaa?ggtactttag?tccagacacc?atggaacgaa

2641?tactctgaca?tgattaagcc?gaagtttaat?ccgtatccaa?atgaaggaga?gaaaattaat

2701?caaatcaaga?aacttcaaaa?actcatagcc?gatgagaaga?gaaaagacac?attagaccga

2761?attaaagtta?acacaattac?taatcctgac?ggagaaaagc?gaatgattgt?taatacgcct

2821?tatggtgttc?aagaaatgtt?cgaggaaact?gaaggaatga?aaagactttc?tcatattgat

2881?ccaaataaga?acgtggctat?tagtgaaact?ccgactgatg?ataacaaaga?atctatttac

2941?agcgccttta?agaacgtttc?accaaatgaa?gcagtcggaa?aattcattga?agatactttt

3001?aattctgcat?actcatccgg?tcaagctcag?agatttgtaa?atcccacaaa?cgcctttact

3061?ggtcaagaag?atccaaagaa?agccagactc?atgactcaga?agactattga?tgttacagag

3121?tattatatta?atgacggtaa?accatcttcg?gctatcaaaa?atcaactaat?ccaaaattac

3181?agagccttag?ctgatagcct?tggaatgacc?gaagaagatt?tcattcaatt?cgctcgtaaa

3241?attcctgatg?attctttggc?acaaatgatc?tcttataatg?aacaatcaga?atcttcgagg

3301?cccgcattag?tcgatgctcc?attctttaag?acacttcagc?cacttgcaaa?tcaaacgtca

3361?aagaccaaac?ttaacgacat?catcaaaata?atcttcacac?aaatttctaa?agtaactggt

3421?aaaaccaatt?caaccacagg?cacaacactc?gagaagaaga?tagttcctaa?tctaaaggca

3481?gtaagatctg?accaagtcat?tgcaggacca?aatggaggaa?cttcagcttt?atcccaagcc

3541?acagctccaa?gaaccggttc?tacagttttg?agtaagcaaa?tcacacgagg?acttcccaga

3601?gttccttcag?gtacaggtac?tagttatcca?gtgagagatc?ccaatggtat?taatacaagt

3661?gaagataatg?aaagatataa?agtcaatcct?tataatccat?attctaaatc?tgatactagt

3721?ggtcttgaag?cacctggtgg?tgaaatagta?cataatggta?caagaccaag?tccttacgct

3781?atagtaggag?ttccaatagt?cgcagcggtt?acaacaccag?tcatcagacc?agcagtctta

3841?agaacgcctc?cagcagcaca?atcgagttct?agtgcacttc?aaggaaatac?agcacttact

3901?ggagcaggaa?caccaagagc?cggagtagca?ggatcaccag?gagtaaatcc?aggaccaact

3961?ggtaaagcac?ctgtctcacc?ccctgcacca?acatcaccag?cttcatctcc?tacagcctca

4021?cctatagcac?ctacagcttc?acctatagca?cctacaggag?catcaatagg?agcacctggt

4081?tcacttggaa?tcccatcatc?accattgaag。

2, a kind of cultivated silkworm microsporidian polar filament protein PTP 3 gene encoded protein matter is characterized in that its sequence is as follows:

1?MIYTLFVIYI?SSTFCRIISR?STVENGPPTV?PFDEHQAKHV?HVRVHGRHRH?GNHHGGGQDV

61?HEVQKHHSHH?HNGHNYGLEH?GKFGNVTYTS?GNHHGHHSHH?GSHADGHHSQ?YGSHADGHHV

121?NAQSDASAPL?DVLPMKEVRS?GDDPALERDL?GIQQAMKAFR?SSLPVSNKDT?NSLLDKAEGI

181?LRSAPDGQLD?APVLDSYGNP?TGQTYREAAE?DAVGYKYPTA?VADEEFDSLV?RSGVEPGRAK

241?QVADKIEETG?FEHEKIKDAV?RNEVARGTAN?RAVAELQDEI?QRRKNATKVK?ERVKVEDELY

301?RQHTLPATGL?FDEDVSVLKN?MSQIVPPAKI?VDYIRDSYDN?SIEAGLPPAV?ARAQADRVGS

361?LITDFNTMLR?SGESPETALS?HLKDMSDRQS?KVDQRRELLQ?NSSPEVLKIL?SDKDNLIKEA

421?LVKTARDPDC?RDRYFDEASQ?KMKILDAREA?SFIHDTENNK?HIRRPRKMAE?DLMDKTNRDN

481?AELVDVAYSH?PVVAKDILKQ?GIANTERVAE?KLADHLDAVK?FAEKAQNPSA?VSALKLLDER

541?KRQDELEKIN?KRNTENEQKM?FKQILKDQED?AEVMQTEEKM?AAANKARKIL?DEGRRAAEQV

601?LKAGGSTDEA?AAVRVAAENA?TARGLEQRES?MIEEEIMKRF?GGAGDPEKAR?KMAAHIAIAH

661?SGENAVGKTP?EEVIKKEQEA?QKKEILRNAL?YDDIGKEPIE?IKSVIPEDRD?EYSIDRSVIK

721?FPLKTFIDEE?VSNVGEAYVK?NKKQSINDEV?RNKVTTTNVN?SGLNVIETEN?GFLNLGENSK

781?KIHIDEATAY?FKPAAKLKME?KKGIKTDNFR?PKTNQGEIRD?MPKGETYYEV?DLKDADLSLP

841?SPTNPTPDTL?VSNGKDVVDV?EDVSAIVINK?GTLVQTPWNE?YSDMIKPKFN?PYPNEGEKIN

901?QIKKLQKLIA?DEKRKDTLDR?IKVNTITNPD?GEKRMIVNTP?YGVQEMFEET?EGMKRLSHID

961?PNKNVAISET?PTDDNKESIY?SAFKNVSPNE?AVGKFIEDTF?NSAYSSGQAQ?RFVNPTNAFT

1021?GQEDPKKARL?MTQKTIDVTE?YYINDGKPSS?AIKNQLIQNY?RALADSLGMT?EEDFIQFARK

1081?IPDDSLAQMI?SYNEQSESSR?PALVDAPFFK?TLQPLANQTS?KTKLNDIIKI?IFTQISKVTG

1141?KTNSTTGTTL?EKKIVPNLKA?VRSDQVIAGP?NGGTSALSQA?TAPRTGSTVL?SKQITRGLPR

1201?VPSGTGTSYP?VRDPNGINTS?EDNERYKVNP?YNPYSKSDTS?GLEAPGGEIV?HNGTRPSPYA

1261?IVGVPIVAAV?TTPVIRPAVL?RTPPAAQSSS?SALQGNTALT?GAGTPRAGVA?GSPGVNPGPT

1321?GKAPVSPPAP?TSPASSPTAS?PIAPTASPIA?PTGASIGAPG?SLGIPSSPLK。

3, the PTP3 gene particularly kills application in the locust agent at the preparation sterilant.