CN112480229B - 蛋白Secp43或其编码基因在调控黑腹果蝇雄性生育中的应用 - Google Patents
蛋白Secp43或其编码基因在调控黑腹果蝇雄性生育中的应用 Download PDFInfo
- Publication number
- CN112480229B CN112480229B CN202011353476.5A CN202011353476A CN112480229B CN 112480229 B CN112480229 B CN 112480229B CN 202011353476 A CN202011353476 A CN 202011353476A CN 112480229 B CN112480229 B CN 112480229B
- Authority
- CN
- China
- Prior art keywords
- secp43
- drosophila melanogaster
- protein
- male sterility
- grna
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 101150071268 TRNAU1AP gene Proteins 0.000 title claims abstract description 44
- 241000255601 Drosophila melanogaster Species 0.000 title claims abstract description 32
- 230000035558 fertility Effects 0.000 title claims abstract description 10
- 108090000623 proteins and genes Proteins 0.000 title abstract description 16
- 230000033228 biological regulation Effects 0.000 title abstract description 8
- 206010021929 Infertility male Diseases 0.000 claims abstract description 15
- 208000007466 Male Infertility Diseases 0.000 claims abstract description 15
- 230000001105 regulatory effect Effects 0.000 claims abstract description 6
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 108020005004 Guide RNA Proteins 0.000 claims description 27
- 239000002773 nucleotide Substances 0.000 claims description 9
- 125000003729 nucleotide group Chemical group 0.000 claims description 9
- 108091033409 CRISPR Proteins 0.000 claims description 8
- 230000001276 controlling effect Effects 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 230000008685 targeting Effects 0.000 claims description 3
- 102000044126 RNA-Binding Proteins Human genes 0.000 abstract description 18
- 102000004169 proteins and genes Human genes 0.000 abstract description 10
- 101710159080 Aconitate hydratase A Proteins 0.000 abstract description 6
- 101710159078 Aconitate hydratase B Proteins 0.000 abstract description 6
- 101710105008 RNA-binding protein Proteins 0.000 abstract description 6
- 208000000509 infertility Diseases 0.000 abstract description 5
- 230000036512 infertility Effects 0.000 abstract description 5
- 231100000535 infertility Toxicity 0.000 abstract description 5
- 210000001625 seminal vesicle Anatomy 0.000 abstract description 4
- 230000001225 therapeutic effect Effects 0.000 abstract description 4
- 201000010653 vesiculitis Diseases 0.000 abstract description 4
- 230000007246 mechanism Effects 0.000 abstract description 3
- 241000255588 Tephritidae Species 0.000 description 15
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 12
- 108700020471 RNA-Binding Proteins Proteins 0.000 description 12
- 239000013598 vector Substances 0.000 description 11
- 239000013612 plasmid Substances 0.000 description 10
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 8
- 238000002347 injection Methods 0.000 description 8
- 239000007924 injection Substances 0.000 description 8
- 238000011161 development Methods 0.000 description 7
- 230000018109 developmental process Effects 0.000 description 7
- 239000012634 fragment Substances 0.000 description 7
- 230000037433 frameshift Effects 0.000 description 7
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 6
- 238000001976 enzyme digestion Methods 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 210000000349 chromosome Anatomy 0.000 description 5
- 230000035772 mutation Effects 0.000 description 5
- 238000012216 screening Methods 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000005520 cutting process Methods 0.000 description 4
- 235000013601 eggs Nutrition 0.000 description 4
- 210000001161 mammalian embryo Anatomy 0.000 description 4
- 239000000178 monomer Substances 0.000 description 4
- 230000009261 transgenic effect Effects 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 3
- 102100037924 Insulin-like growth factor 2 mRNA-binding protein 1 Human genes 0.000 description 3
- 101710126181 Insulin-like growth factor 2 mRNA-binding protein 1 Proteins 0.000 description 3
- 102100040347 TAR DNA-binding protein 43 Human genes 0.000 description 3
- 101710150875 TAR DNA-binding protein 43 Proteins 0.000 description 3
- 238000012217 deletion Methods 0.000 description 3
- 230000037430 deletion Effects 0.000 description 3
- 210000002257 embryonic structure Anatomy 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 230000014509 gene expression Effects 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 238000000520 microinjection Methods 0.000 description 3
- 230000004770 neurodegeneration Effects 0.000 description 3
- 208000015122 neurodegenerative disease Diseases 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 208000005623 Carcinogenesis Diseases 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- PKVWNYGXMNWJSI-CIUDSAMLSA-N Gln-Gln-Gln Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O PKVWNYGXMNWJSI-CIUDSAMLSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- QBHGXFQJFPWJIH-XUXIUFHCSA-N Lys-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CCCCN QBHGXFQJFPWJIH-XUXIUFHCSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 102000004389 Ribonucleoproteins Human genes 0.000 description 2
- 108010081734 Ribonucleoproteins Proteins 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 230000036952 cancer formation Effects 0.000 description 2
- 231100000504 carcinogenesis Toxicity 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 239000006059 cover glass Substances 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 108010040030 histidinoalanine Proteins 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000013011 mating Effects 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 231100001160 nonlethal Toxicity 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000001131 transforming effect Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 102000007469 Actins Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- AAQGRPOPTAUUBM-ZLUOBGJFSA-N Ala-Ala-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O AAQGRPOPTAUUBM-ZLUOBGJFSA-N 0.000 description 1
- UGLPMYSCWHTZQU-AUTRQRHGSA-N Ala-Ala-Tyr Chemical compound C[C@H]([NH3+])C(=O)N[C@@H](C)C(=O)N[C@H](C([O-])=O)CC1=CC=C(O)C=C1 UGLPMYSCWHTZQU-AUTRQRHGSA-N 0.000 description 1
- PBAMJJXWDQXOJA-FXQIFTODSA-N Ala-Asp-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N PBAMJJXWDQXOJA-FXQIFTODSA-N 0.000 description 1
- OQCPATDFWYYDDX-HGNGGELXSA-N Ala-Gln-His Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O OQCPATDFWYYDDX-HGNGGELXSA-N 0.000 description 1
- FUSPCLTUKXQREV-ACZMJKKPSA-N Ala-Glu-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O FUSPCLTUKXQREV-ACZMJKKPSA-N 0.000 description 1
- KXEVYGKATAMXJJ-ACZMJKKPSA-N Ala-Glu-Asp Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O KXEVYGKATAMXJJ-ACZMJKKPSA-N 0.000 description 1
- ZVFVBBGVOILKPO-WHFBIAKZSA-N Ala-Gly-Ala Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O ZVFVBBGVOILKPO-WHFBIAKZSA-N 0.000 description 1
- VGPWRRFOPXVGOH-BYPYZUCNSA-N Ala-Gly-Gly Chemical compound C[C@H](N)C(=O)NCC(=O)NCC(O)=O VGPWRRFOPXVGOH-BYPYZUCNSA-N 0.000 description 1
- HQJKCXHQNUCKMY-GHCJXIJMSA-N Ala-Ile-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](C)N HQJKCXHQNUCKMY-GHCJXIJMSA-N 0.000 description 1
- LBYMZCVBOKYZNS-CIUDSAMLSA-N Ala-Leu-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O LBYMZCVBOKYZNS-CIUDSAMLSA-N 0.000 description 1
- CCDFBRZVTDDJNM-GUBZILKMSA-N Ala-Leu-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O CCDFBRZVTDDJNM-GUBZILKMSA-N 0.000 description 1
- RGQCNKIDEQJEBT-CQDKDKBSSA-N Ala-Leu-Tyr Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 RGQCNKIDEQJEBT-CQDKDKBSSA-N 0.000 description 1
- MDNAVFBZPROEHO-UHFFFAOYSA-N Ala-Lys-Val Natural products CC(C)C(C(O)=O)NC(=O)C(NC(=O)C(C)N)CCCCN MDNAVFBZPROEHO-UHFFFAOYSA-N 0.000 description 1
- AWNAEZICPNGAJK-FXQIFTODSA-N Ala-Met-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CO)C(O)=O AWNAEZICPNGAJK-FXQIFTODSA-N 0.000 description 1
- KLALXKYLOMZDQT-ZLUOBGJFSA-N Ala-Ser-Asn Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC(N)=O KLALXKYLOMZDQT-ZLUOBGJFSA-N 0.000 description 1
- NLYYHIKRBRMAJV-AEJSXWLSSA-N Ala-Val-Pro Chemical compound C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N NLYYHIKRBRMAJV-AEJSXWLSSA-N 0.000 description 1
- XEPSCVXTCUUHDT-AVGNSLFASA-N Arg-Arg-Leu Natural products CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CCCN=C(N)N XEPSCVXTCUUHDT-AVGNSLFASA-N 0.000 description 1
- PZBSKYJGKNNYNK-ULQDDVLXSA-N Arg-Leu-Tyr Chemical compound CC(C)C[C@H](NC(=O)[C@@H](N)CCCN=C(N)N)C(=O)N[C@@H](Cc1ccc(O)cc1)C(O)=O PZBSKYJGKNNYNK-ULQDDVLXSA-N 0.000 description 1
- INXWADWANGLMPJ-JYJNAYRXSA-N Arg-Phe-Arg Chemical compound NC(=N)NCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)CC1=CC=CC=C1 INXWADWANGLMPJ-JYJNAYRXSA-N 0.000 description 1
- BDMIFVIWCNLDCT-CIUDSAMLSA-N Asn-Arg-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O BDMIFVIWCNLDCT-CIUDSAMLSA-N 0.000 description 1
- NTWOPSIUJBMNRI-KKUMJFAQSA-N Asn-Lys-Tyr Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 NTWOPSIUJBMNRI-KKUMJFAQSA-N 0.000 description 1
- MVXJBVVLACEGCG-PCBIJLKTSA-N Asn-Phe-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O MVXJBVVLACEGCG-PCBIJLKTSA-N 0.000 description 1
- NJSNXIOKBHPFMB-GMOBBJLQSA-N Asn-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CC(=O)N)N NJSNXIOKBHPFMB-GMOBBJLQSA-N 0.000 description 1
- QYRMBFWDSFGSFC-OLHMAJIHSA-N Asn-Thr-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O QYRMBFWDSFGSFC-OLHMAJIHSA-N 0.000 description 1
- ZCKYZTGLXIEOKS-CIUDSAMLSA-N Asp-Asp-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)O)N ZCKYZTGLXIEOKS-CIUDSAMLSA-N 0.000 description 1
- UMHUHHJMEXNSIV-CIUDSAMLSA-N Asp-Leu-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(O)=O UMHUHHJMEXNSIV-CIUDSAMLSA-N 0.000 description 1
- GKWFMNNNYZHJHV-SRVKXCTJSA-N Asp-Lys-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC(O)=O GKWFMNNNYZHJHV-SRVKXCTJSA-N 0.000 description 1
- RVMXMLSYBTXCAV-VEVYYDQMSA-N Asp-Pro-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O RVMXMLSYBTXCAV-VEVYYDQMSA-N 0.000 description 1
- QSFHZPQUAAQHAQ-CIUDSAMLSA-N Asp-Ser-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O QSFHZPQUAAQHAQ-CIUDSAMLSA-N 0.000 description 1
- OYSYWMMZGJSQRB-AVGNSLFASA-N Asp-Tyr-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O OYSYWMMZGJSQRB-AVGNSLFASA-N 0.000 description 1
- CZIVKMOEXPILDK-SRVKXCTJSA-N Asp-Tyr-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(O)=O CZIVKMOEXPILDK-SRVKXCTJSA-N 0.000 description 1
- 241001136675 Buddleja davidii Species 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- OVQXQLWWJSNYFV-XEGUGMAKSA-N Gln-Ala-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CCC(N)=O)C)C(O)=O)=CNC2=C1 OVQXQLWWJSNYFV-XEGUGMAKSA-N 0.000 description 1
- JESJDAAGXULQOP-CIUDSAMLSA-N Gln-Arg-Ser Chemical compound C(C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)CN=C(N)N JESJDAAGXULQOP-CIUDSAMLSA-N 0.000 description 1
- YXQCLIVLWCKCRS-RYUDHWBXSA-N Gln-Gly-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCC(=O)N)N)O YXQCLIVLWCKCRS-RYUDHWBXSA-N 0.000 description 1
- QDXMSSWCEVYOLZ-SZMVWBNQSA-N Gln-Leu-Trp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CCC(=O)N)N QDXMSSWCEVYOLZ-SZMVWBNQSA-N 0.000 description 1
- ZEEPYMXTJWIMSN-GUBZILKMSA-N Gln-Lys-Ser Chemical compound NCCCC[C@@H](C(=O)N[C@@H](CO)C(O)=O)NC(=O)[C@@H](N)CCC(N)=O ZEEPYMXTJWIMSN-GUBZILKMSA-N 0.000 description 1
- KVQOVQVGVKDZNW-GUBZILKMSA-N Gln-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)N)N KVQOVQVGVKDZNW-GUBZILKMSA-N 0.000 description 1
- NHMRJKKAVMENKJ-WDCWCFNPSA-N Gln-Thr-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O NHMRJKKAVMENKJ-WDCWCFNPSA-N 0.000 description 1
- WIMVKDYAKRAUCG-IHRRRGAJSA-N Gln-Tyr-Glu Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O WIMVKDYAKRAUCG-IHRRRGAJSA-N 0.000 description 1
- HPBKQFJXDUVNQV-FHWLQOOXSA-N Gln-Tyr-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O HPBKQFJXDUVNQV-FHWLQOOXSA-N 0.000 description 1
- CGYDXNKRIMJMLV-GUBZILKMSA-N Glu-Arg-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O CGYDXNKRIMJMLV-GUBZILKMSA-N 0.000 description 1
- CKRUHITYRFNUKW-WDSKDSINSA-N Glu-Asn-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O CKRUHITYRFNUKW-WDSKDSINSA-N 0.000 description 1
- XXCDTYBVGMPIOA-FXQIFTODSA-N Glu-Asp-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O XXCDTYBVGMPIOA-FXQIFTODSA-N 0.000 description 1
- CCQOOWAONKGYKQ-BYPYZUCNSA-N Gly-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)CN CCQOOWAONKGYKQ-BYPYZUCNSA-N 0.000 description 1
- GGAPHLIUUTVYMX-QWRGUYRKSA-N Gly-Phe-Ser Chemical compound OC[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)C[NH3+])CC1=CC=CC=C1 GGAPHLIUUTVYMX-QWRGUYRKSA-N 0.000 description 1
- SOEGEPHNZOISMT-BYPYZUCNSA-N Gly-Ser-Gly Chemical compound NCC(=O)N[C@@H](CO)C(=O)NCC(O)=O SOEGEPHNZOISMT-BYPYZUCNSA-N 0.000 description 1
- WRFOZIJRODPLIA-QWRGUYRKSA-N Gly-Tyr-Cys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)CN)O WRFOZIJRODPLIA-QWRGUYRKSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- RVKIPWVMZANZLI-UHFFFAOYSA-N H-Lys-Trp-OH Natural products C1=CC=C2C(CC(NC(=O)C(N)CCCCN)C(O)=O)=CNC2=C1 RVKIPWVMZANZLI-UHFFFAOYSA-N 0.000 description 1
- 102000006479 Heterogeneous-Nuclear Ribonucleoproteins Human genes 0.000 description 1
- 108010019372 Heterogeneous-Nuclear Ribonucleoproteins Proteins 0.000 description 1
- IGBBXBFSLKRHJB-BZSNNMDCSA-N His-Lys-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CN=CN1 IGBBXBFSLKRHJB-BZSNNMDCSA-N 0.000 description 1
- NKVZTQVGUNLLQW-JBDRJPRFSA-N Ile-Ala-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)O)N NKVZTQVGUNLLQW-JBDRJPRFSA-N 0.000 description 1
- LGMUPVWZEYYUMU-YVNDNENWSA-N Ile-Glu-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N LGMUPVWZEYYUMU-YVNDNENWSA-N 0.000 description 1
- LZDNBBYBDGBADK-UHFFFAOYSA-N L-valyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C(C)C)C(O)=O)=CNC2=C1 LZDNBBYBDGBADK-UHFFFAOYSA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- OIARJGNVARWKFP-YUMQZZPRSA-N Leu-Asn-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O OIARJGNVARWKFP-YUMQZZPRSA-N 0.000 description 1
- OGCQGUIWMSBHRZ-CIUDSAMLSA-N Leu-Asn-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O OGCQGUIWMSBHRZ-CIUDSAMLSA-N 0.000 description 1
- WQWSMEOYXJTFRU-GUBZILKMSA-N Leu-Glu-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O WQWSMEOYXJTFRU-GUBZILKMSA-N 0.000 description 1
- HVJVUYQWFYMGJS-GVXVVHGQSA-N Leu-Glu-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O HVJVUYQWFYMGJS-GVXVVHGQSA-N 0.000 description 1
- HYMLKESRWLZDBR-WEDXCCLWSA-N Leu-Gly-Thr Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O HYMLKESRWLZDBR-WEDXCCLWSA-N 0.000 description 1
- PKKMDPNFGULLNQ-AVGNSLFASA-N Leu-Met-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O PKKMDPNFGULLNQ-AVGNSLFASA-N 0.000 description 1
- UCBPDSYUVAAHCD-UWVGGRQHSA-N Leu-Pro-Gly Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O UCBPDSYUVAAHCD-UWVGGRQHSA-N 0.000 description 1
- BTEMNFBEAAOGBR-BZSNNMDCSA-N Leu-Tyr-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BTEMNFBEAAOGBR-BZSNNMDCSA-N 0.000 description 1
- AIMGJYMCTAABEN-GVXVVHGQSA-N Leu-Val-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O AIMGJYMCTAABEN-GVXVVHGQSA-N 0.000 description 1
- 108020005198 Long Noncoding RNA Proteins 0.000 description 1
- NNKLKUUGESXCBS-KBPBESRZSA-N Lys-Gly-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O NNKLKUUGESXCBS-KBPBESRZSA-N 0.000 description 1
- XDPLZVNMYQOFQZ-BJDJZHNGSA-N Lys-Ile-Cys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCCCN)N XDPLZVNMYQOFQZ-BJDJZHNGSA-N 0.000 description 1
- LUTDBHBIHHREDC-IHRRRGAJSA-N Lys-Pro-Lys Chemical compound NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(O)=O LUTDBHBIHHREDC-IHRRRGAJSA-N 0.000 description 1
- WXHHTBVYQOSYSL-FXQIFTODSA-N Met-Ala-Ser Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O WXHHTBVYQOSYSL-FXQIFTODSA-N 0.000 description 1
- ACYHZNZHIZWLQF-BQBZGAKWSA-N Met-Asn-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O ACYHZNZHIZWLQF-BQBZGAKWSA-N 0.000 description 1
- YCUSPBPZVJDMII-YUMQZZPRSA-N Met-Gly-Glu Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O YCUSPBPZVJDMII-YUMQZZPRSA-N 0.000 description 1
- LRALLISKBZNSKN-BQBZGAKWSA-N Met-Gly-Ser Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O LRALLISKBZNSKN-BQBZGAKWSA-N 0.000 description 1
- DYTWOWJWJCBFLE-IHRRRGAJSA-N Met-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CCSC)CC1=CNC=N1 DYTWOWJWJCBFLE-IHRRRGAJSA-N 0.000 description 1
- GMMLGMFBYCFCCX-KZVJFYERSA-N Met-Thr-Ala Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O GMMLGMFBYCFCCX-KZVJFYERSA-N 0.000 description 1
- FXBKQTOGURNXSL-HJGDQZAQSA-N Met-Thr-Glu Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCC(O)=O FXBKQTOGURNXSL-HJGDQZAQSA-N 0.000 description 1
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 1
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 1
- WUGMRIBZSVSJNP-UHFFFAOYSA-N N-L-alanyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C)C(O)=O)=CNC2=C1 WUGMRIBZSVSJNP-UHFFFAOYSA-N 0.000 description 1
- XZFYRXDAULDNFX-UHFFFAOYSA-N N-L-cysteinyl-L-phenylalanine Natural products SCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XZFYRXDAULDNFX-UHFFFAOYSA-N 0.000 description 1
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 1
- 108010066427 N-valyltryptophan Proteins 0.000 description 1
- 108010087066 N2-tryptophyllysine Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- YYRCPTVAPLQRNC-ULQDDVLXSA-N Phe-Arg-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](N)CC1=CC=CC=C1 YYRCPTVAPLQRNC-ULQDDVLXSA-N 0.000 description 1
- HGNGAMWHGGANAU-WHOFXGATSA-N Phe-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=CC=C1 HGNGAMWHGGANAU-WHOFXGATSA-N 0.000 description 1
- CWFGECHCRMGPPT-MXAVVETBSA-N Phe-Ile-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O CWFGECHCRMGPPT-MXAVVETBSA-N 0.000 description 1
- MVIJMIZJPHQGEN-IHRRRGAJSA-N Phe-Ser-Val Chemical compound CC(C)[C@@H](C([O-])=O)NC(=O)[C@H](CO)NC(=O)[C@@H]([NH3+])CC1=CC=CC=C1 MVIJMIZJPHQGEN-IHRRRGAJSA-N 0.000 description 1
- GLUYKHMBGKQBHE-JYJNAYRXSA-N Phe-Val-Arg Chemical compound NC(=N)NCCC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 GLUYKHMBGKQBHE-JYJNAYRXSA-N 0.000 description 1
- GOUWCZRDTWTODO-YDHLFZDLSA-N Phe-Val-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O GOUWCZRDTWTODO-YDHLFZDLSA-N 0.000 description 1
- AFXCXDQNRXTSBD-FJXKBIBVSA-N Pro-Gly-Thr Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O AFXCXDQNRXTSBD-FJXKBIBVSA-N 0.000 description 1
- GZNYIXWOIUFLGO-ZJDVBMNYSA-N Pro-Thr-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZNYIXWOIUFLGO-ZJDVBMNYSA-N 0.000 description 1
- 101000702488 Rattus norvegicus High affinity cationic amino acid transporter 1 Proteins 0.000 description 1
- DKKGAAJTDKHWOD-BIIVOSGPSA-N Ser-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CO)N)C(=O)O DKKGAAJTDKHWOD-BIIVOSGPSA-N 0.000 description 1
- SWSRFJZZMNLMLY-ZKWXMUAHSA-N Ser-Asp-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O SWSRFJZZMNLMLY-ZKWXMUAHSA-N 0.000 description 1
- LALNXSXEYFUUDD-GUBZILKMSA-N Ser-Glu-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LALNXSXEYFUUDD-GUBZILKMSA-N 0.000 description 1
- ZOPISOXXPQNOCO-SVSWQMSJSA-N Ser-Ile-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](CO)N ZOPISOXXPQNOCO-SVSWQMSJSA-N 0.000 description 1
- WGDYNRCOQRERLZ-KKUMJFAQSA-N Ser-Lys-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)N WGDYNRCOQRERLZ-KKUMJFAQSA-N 0.000 description 1
- OZPDGESCTGGNAD-CIUDSAMLSA-N Ser-Ser-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CO OZPDGESCTGGNAD-CIUDSAMLSA-N 0.000 description 1
- ZKOKTQPHFMRSJP-YJRXYDGGSA-N Ser-Thr-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZKOKTQPHFMRSJP-YJRXYDGGSA-N 0.000 description 1
- HKHCTNFKZXAMIF-KKUMJFAQSA-N Ser-Tyr-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)CC1=CC=C(O)C=C1 HKHCTNFKZXAMIF-KKUMJFAQSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- ZUXQFMVPAYGPFJ-JXUBOQSCSA-N Thr-Ala-Lys Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCCN ZUXQFMVPAYGPFJ-JXUBOQSCSA-N 0.000 description 1
- DWYAUVCQDTZIJI-VZFHVOOUSA-N Thr-Ala-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O DWYAUVCQDTZIJI-VZFHVOOUSA-N 0.000 description 1
- AQAMPXBRJJWPNI-JHEQGTHGSA-N Thr-Gly-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O AQAMPXBRJJWPNI-JHEQGTHGSA-N 0.000 description 1
- NQQMWWVVGIXUOX-SVSWQMSJSA-N Thr-Ser-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O NQQMWWVVGIXUOX-SVSWQMSJSA-N 0.000 description 1
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 1
- 108700019146 Transgenes Proteins 0.000 description 1
- WPSYJHFHZYJXMW-JSGCOSHPSA-N Trp-Gln-Gly Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O WPSYJHFHZYJXMW-JSGCOSHPSA-N 0.000 description 1
- GWBWCGITOYODER-YTQUADARSA-N Trp-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N GWBWCGITOYODER-YTQUADARSA-N 0.000 description 1
- SUEGAFMNTXXNLR-WFBYXXMGSA-N Trp-Ser-Ala Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O SUEGAFMNTXXNLR-WFBYXXMGSA-N 0.000 description 1
- XQMGDVVKFRLQKH-BBRMVZONSA-N Trp-Val-Gly Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O)=CNC2=C1 XQMGDVVKFRLQKH-BBRMVZONSA-N 0.000 description 1
- QUILOGWWLXMSAT-IHRRRGAJSA-N Tyr-Gln-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O QUILOGWWLXMSAT-IHRRRGAJSA-N 0.000 description 1
- WAPFQMXRSDEGOE-IHRRRGAJSA-N Tyr-Glu-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O WAPFQMXRSDEGOE-IHRRRGAJSA-N 0.000 description 1
- NMKJPMCEKQHRPD-IRXDYDNUSA-N Tyr-Gly-Tyr Chemical compound C([C@H](N)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 NMKJPMCEKQHRPD-IRXDYDNUSA-N 0.000 description 1
- GGXUDPQWAWRINY-XEGUGMAKSA-N Tyr-Ile-Gly Chemical compound OC(=O)CNC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 GGXUDPQWAWRINY-XEGUGMAKSA-N 0.000 description 1
- WFENBJPLZMPVAX-XVKPBYJWSA-N Val-Gly-Glu Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O WFENBJPLZMPVAX-XVKPBYJWSA-N 0.000 description 1
- MANXHLOVEUHVFD-DCAQKATOSA-N Val-His-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CS)C(=O)O)N MANXHLOVEUHVFD-DCAQKATOSA-N 0.000 description 1
- FTKXYXACXYOHND-XUXIUFHCSA-N Val-Ile-Leu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O FTKXYXACXYOHND-XUXIUFHCSA-N 0.000 description 1
- AGXGCFSECFQMKB-NHCYSSNCSA-N Val-Leu-Asp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N AGXGCFSECFQMKB-NHCYSSNCSA-N 0.000 description 1
- KISFXYYRKKNLOP-IHRRRGAJSA-N Val-Phe-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)O)N KISFXYYRKKNLOP-IHRRRGAJSA-N 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 108010008685 alanyl-glutamyl-aspartic acid Proteins 0.000 description 1
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 1
- 108010044940 alanylglutamine Proteins 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 210000003484 anatomy Anatomy 0.000 description 1
- 108010080488 arginyl-arginyl-leucine Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 230000032669 eclosion Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000003754 fetus Anatomy 0.000 description 1
- 238000002194 freeze distillation Methods 0.000 description 1
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 108010073628 glutamyl-valyl-phenylalanine Proteins 0.000 description 1
- 108010045126 glycyl-tyrosyl-glycine Proteins 0.000 description 1
- 108010015792 glycyllysine Proteins 0.000 description 1
- 108010087823 glycyltyrosine Proteins 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 150000008282 halocarbons Chemical class 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 102000028416 insulin-like growth factor binding Human genes 0.000 description 1
- 108091022911 insulin-like growth factor binding Proteins 0.000 description 1
- 108010083708 leucyl-aspartyl-valine Proteins 0.000 description 1
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 1
- 108010051673 leucyl-glycyl-phenylalanine Proteins 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000021121 meiosis Effects 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 108010016686 methionyl-alanyl-serine Proteins 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000004879 molecular function Effects 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000011837 pasties Nutrition 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000001124 posttranscriptional effect Effects 0.000 description 1
- 230000007859 posttranscriptional regulation of gene expression Effects 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000004153 renaturation Methods 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000005740 tumor formation Effects 0.000 description 1
- 108010003137 tyrosyltyrosine Proteins 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
- A01K67/0333—Genetically modified invertebrates, e.g. transgenic, polyploid
- A01K67/0337—Genetically modified Arthropods
- A01K67/0339—Genetically modified insects, e.g. Drosophila melanogaster, medfly
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/07—Animals genetically altered by homologous recombination
- A01K2217/075—Animals genetically altered by homologous recombination inducing loss of function, i.e. knock out
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/70—Invertebrates
- A01K2227/706—Insects, e.g. Drosophila melanogaster, medfly
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/20—Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPRs]
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Environmental Sciences (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Animal Behavior & Ethology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了蛋白Secp43或其编码基因在调控黑腹果蝇雄性生育中的应用。所述的蛋白Secp43，其氨基酸序列如SEQ ID NO.2所示。本发明通过发现，敲除编码基因Secp43可以使得黑腹果蝇雄性不育，其成束精子无法完成分离，未成熟的精子则无法进入储精囊，这可能是雄性不育的主要原因。因此本发明筛选得到一个黑腹果蝇雄性不育相关的新蛋白，鉴于果蝇和人类部分蛋白的同源性，将有助于我们寻找到新的不孕不育的治疗靶点。并且从RNA结合蛋白角度对Secp43造成黑腹果蝇雄性不育的调控机理进行深入研究，将有助于我们寻找到新的不孕不育的治疗靶标。
Description
技术领域：
本发明属于生物技术领域，具体地涉及蛋白Secp43或其编码基因在调控黑腹果蝇雄性生育中的应用。
背景技术：
在真核生物中,基因表达的转录后调节是一个非常复杂和关键的步骤,在RNA代谢途径中，都需要大量蛋白质因子相互作用，同时RNA代谢还受到多种信号途径的调节。根据RNA的功能和位置可以把RNA分为很多种类，每一种RNA都有大量RNA结合蛋白和它相互作用来稳定、保护、组装或者转移它，同时还调节RNA同其它分子的相互作用或者催化RNA解螺旋和复制等。RNA结合蛋白的研究，历来受到相关生物化学家们的重视，已经有很多不同结构和功能的RNA结合蛋白已经被鉴定出来。目前研究得比较清楚的RNA结合蛋白有核糖核蛋白(RNP)、KH基序RNA结合蛋白、双螺旋RNA结合蛋白和冷激基序RNA结合蛋白(Graumann andMarahiel,1998)。然而，还有大量RNA结合蛋白的功能尚未清楚。
近年来,对动植物中RNA结合蛋白的研究有很多,但对RNA结合蛋白的生理功能和分子功能研究还有待深入。RNA结合蛋白和动物的神经系统发育密切相关。目前，RNA结合蛋白在神经退行性疾病的发生中所起到的作用受到广泛关注。在2006年和2009年，TDP-43和FUS相继被报道与渐冻人症和其他几种神经退行性疾病的发生密切相关(Kwiatkowski etal.,2009；Neumann et al.,2006)。TDP-43和FUS都是RNA结合蛋白，具有RRM结构域和甘氨酸富集结构域(glycine rich domain,GRD)，均属于hnRNP家族。TDP-43和FUS作为RNA结合蛋白在RNA转录后调控上发挥着重要作用，大量证据表明它们的功能异常可能与神经退行性疾病的发生紧密相关。
此外,在肿瘤发生和调节机制上对RNA结合蛋白的研究也很多。Charles等对CRD-BP进行研究发现，CRD-BP是一种多功能的RNA结合蛋白，它结合c-myc、肌动蛋白mRNAs、胰岛素样生长因子结合蛋白ⅡmRNAs和H19 RNA(Liao et al.,2004)。CRD-BP在胎儿中含量很高但在成人中很低或者没有，他们在一些成年人的乳房、结肠和肺部中检测到CRD-BP的表达，且这这些部位产生癌变的概率很高。因此他们推测CRD-BP涉及肿瘤的形成。
由于生殖发育一直是人类医学关注的重点之一，其不孕不育的治疗靶点应该多种多样。
发明内容：
本发明的第一个目的是提供蛋白Secp43或其编码基因Secp43在调控黑腹果蝇雄性生育中的应用，所述的蛋白Secp43，其氨基酸序列如SEQ ID NO.2所示。
优选，所述的蛋白Secp43的编码基因Secp43，其核苷酸序列如SEQ ID NO.1所示。
优选，敲除编码基因Secp43在调控黑腹果蝇雄性不育中的应用。
所述的敲除编码基因Secp43优选是利用Crispr/Cas9特异性敲除编码基因Secp43，其中用于靶向编码基因Secp43的gRNA，其核苷酸序列如SEQ ID NO.3所示。
本发明的第二个目的是提供一种在利用Crispr/Cas9特异性敲除编码基因Secp43中用于靶向编码基因Secp43的gRNA，其核苷酸序列如SEQ ID NO.3所示。
本发明的第三个目的是提供上述gRNA在制备敲除编码基因Secp43从而调控黑腹果蝇雄性不育的制剂中的应用。
本发明通过发现，敲除编码基因Secp43可以使得黑腹果蝇雄性不育，其成束精子无法完成分离，未成熟的精子则无法进入储精囊，这可能是雄性不育的主要原因。因此本发明筛选得到一个黑腹果蝇雄性不育相关的新蛋白，鉴于果蝇和人类部分蛋白的同源性，将有助于我们寻找到新的不孕不育的治疗靶点。并且从RNA结合蛋白角度对Secp43造成黑腹果蝇雄性不育的调控机理进行深入研究，将有助于我们寻找到新的不孕不育的治疗靶标。
附图说明：
图1是Secp43的杂合突变体检测图。箭头表示突变体。NC表示野生型果蝇经过PCR扩增与酶切之后的结果；
图2是黑腹果蝇的生育力统计。每个点表示平均数±SEM(n＝30)，字母表示多重分析结果(P＜0.05，统计方法为LSR，SPSS)；
图3是敲除Secp43后黑腹果蝇雄性成虫精巢的解剖图。
具体实施方式：
下面结合实施例及附图对本发明作进一步详细的描述，但本发明的实施方式不限于此。
实施例1
1.目标基因，其核苷酸序列如下所示：
atggcgtccgtgcattgtcaactgtggatgggaagcctggagtcctacatgacggagaacttcataatcgccgctttccggaagatgggcgaagatcccaccacggtgcgcctgatgcgcaacaagtacacgggcgaaccggccggctactgcttcgtcaacttcatatccgatgaccatgcgctggacgcgatgcacaagctaaacggtaagcccattccgggcaccaatcccattgtgcgattccgtcttaactcggccagcaactcgtacaagctgcccgggaatgaacgcgaatttagcgtctgggtgggcgacctcagctcggacgtggacgactatcagctgtacaaggtgttctccagcaagttcacatcgatcaaaaccgccaaggtgatcttggacagcctggggttttccaagggttacggctttgtgcgatttggcatcgaggatgagcagaaatcagcgctgtacgacatgaacgggtacatcggattgggtaccaagcccataaagatttgcaatgccgtgcccaaacccaaatccgagttgggtggtgctgtgggcgaaggcaacacaaactatggatatggaagcggtatgactgcagcaggtggcaccgactactcgcagtactacgaccccaccagcacctattggcagggataccaggcctggcaaggttactacgagcaggccggtgcttcgatcacagatgcggctgcatactaccagcaggccatgtcgcagtcgcactctaatccgcagactctggcccagcacgccgaggcctggagcgcacaacgcagcgcccagtacgagcagcagcagcagcagcagaccgcttccgccgccaatggagctgaggatgagaacggactggtggagcacaagttcgtcctggatgtggacaaactgaaccgagaggccattgatgccgaccgccgtctgtacgatgctctggagagctctaagtggctgcccattgagcagctggaagtcttttag(具体序列如SEQ ID NO.1所示)，其编码的蛋白的氨基酸序列如SEQ ID NO.2所示，命名为蛋白Secp43。
登陆Crispr在线gRNA靶标网站选取合适的gRNA，http:// tools.flycrispr.molbio.wisc.edu/targetFinder/#userconsent#，蛋白通常只需1个gRNA即可产生失效蛋白。设计时，在gRNA的两端各加上了bbs I酶切后的序列。此外，为了便于快速检测突变体，我们选择target部位含有常用酶切位点gRNA序列，如发生突变，酶切则失效，反之则反。我们为Secp43选择了1个gRNA(靶点)，序列如下：
gRNA：5’-ATGGGCGAAGATCCCACCA-3’，其核苷酸序列如SEQ ID NO.3所示。
2.gRNA单体质粒的构建：包括短引物替换法构建gRNA单体、pMD18T-U6-Bbs I载体酶切、载体与gRNA片段的连接、转化及重组质粒的筛选鉴定。具体如下：
(1)短引物替换法构建gRNA单体。
1)将合成的引物(FP:GTCGATGGGCGAAGATCCCACCA；RP:AAACTGGTGGGATCTTCGCCCAT)成对摆放好,13000g离心2min。
2)加入TE Buffer将引物稀释至100μM。稀释完毕后各取2μl混合,成为终浓度为10μM的引物。
3)94℃高温处理5min使引物复性结合，冰上备用。
(2)酶切载体(pMD18T-U6-Bbs I)
将反应物混匀后稍离心，37℃水浴中酶切2-4h。
(3)载体与gRNA片段的连接、转化及重组质粒的筛选鉴定
将gRNA目的片段和pMD18-T-U6载体利用T4 DNA连接酶进行体外连接，反应体系如下：
将反应物混匀后，稍离心，16℃连接2h。然后将连接产物转化进表达菌大肠杆菌DH5a中，通过PCR鉴定gRNA是否插入载体，并进行测序验证，将鉴定正确的菌液制成甘油菌，-80℃冰箱保存。
所述的pMD18-T-U6载体是在pMD18-T中的bbs I酶切位插入以下序列：
5'-GCTCACCTGTGATTGCTCCTACTCAAATACAAAAACATCAAATTTTCTGTCAATAAAGCATATTTATTTATATTTATTTTACAGGAAAGAATTCCTTTTAAAGTGTATTTTAACCTATAATGAAAAACGATTAAAAAAAATACATAAAATAATTCGAAAATTTTTGAATAGCCCAGGTTGATAAAAATTCATTTCATACGTTTTATAACTTATGCCCCTAAGTATTTTTTGACCATAGTGTTTCAATTCTACATTAATTTTACAGAGTAGAATGAAACGCCACCTACTCAGCCAAGAGGCGAAAAGGTTAGCTCGCCAAGCAGAGAGGGCGCCAGTGCTCACTACTTTTTATAATTCTCAACTTCTTTTTCCAGACTCAGTTCGTATATATAGACCTATTTTCAATTTAACGTCGGGGTCTTCGAGAAGACCTGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTGCCTACCTGGAGCCTGAGAGTTGTTCAATAAAATAAAAATGTTTCGTTTTTTTGCTTTCGCCAGTATTTATTATTTTTCATCAATATGTATTCAATTTGGTATGTATTTAGTAATTGTAATATATAGACAATGGTTTTCCGTTGACG-3'
3.PUAST-attB注射载体的构建：包括将单体质粒(步骤2最后构建的保存在甘油菌中的含gRNA片段的重组质粒)亚克隆到Gateway载体中和转化及重组质粒的筛选鉴定。具体如下：
(1)将单体质粒亚克隆到Gateway载体中
将反应物混匀后稍离心，室温反应1-2h。
(2)转化及重组质粒的筛选鉴定
将反应产物加入1ul蛋白酶K于37℃水浴10min，转化进表达菌大肠杆菌DH5a中，通过EcoR I和BamH I酶切鉴定3个gRNA是否插入载体，将鉴定正确的菌液制成甘油菌，-80℃冰箱保存，提取其中重组质粒可用于注射。
4.突变体果蝇的制备及杂交。
(1)注射前胚胎的收集和处理
①在大培养瓶中收集300只左右的待注射果蝇，保证雌果蝇与雄果蝇比例在2:1，果蝇年龄控制在3天以内，让其交配充分。
②交配2天之后，开始收集胚胎。收集时将果蝇倒入收卵器中，收集卵的果汁平皿涂上一层糊状的酵母。将瓶子倒置放在安静的环境中，25℃静置。
③每隔1-2h即可更新果汁平皿，将收集到的卵在显微镜下剥壳，按统一方向排列整齐后，粘到带有双面胶的盖玻片上，在硅胶中干燥2-3分钟，以减少卵的内压。干燥以后，用一薄层的700卤化碳油盖好。
(2)注射针头的制备
用拉针仪将显微注射用的玻璃毛细针管加热拉制针头。在针头中灌入待注射溶液，将针头在显微镜盖玻片的周围轻蹭一下，以此折断针头保证液体流出。
(3)显微注射操作
显微注射在18℃(以减缓发育的速度)、相对湿度大于70％的房间内完成。将带有胚胎的载玻片置于倒置显微镜下，注射的针头入针点可定在胚胎长的5％-10％处，可以移动显微操作载物台来调节位置。在胚胎的尾部前三分之一进行注射(注射的是上述含有gRNA的重组质粒，浓度500ng/ul，体积大约1-2nl)。注射结束后，将带有胚胎的盖玻片放到湿润的培养皿中，置于18摄氏度的培养箱中使其继续发育。
(4)转基因果蝇的饲养和筛选
注射后24-36小时，胚胎即可发育为1龄幼虫，将这些幼虫用毛刷轻挑至带有酵母的玉米培养基中，约50只幼虫一瓶。十天以后，待果蝇羽化后尽快挑出避免其互相交配，并使每一只羽化果蝇与野生型w1118果蝇杂交，w1118的雌果蝇应挑选处女蝇。约两周后，检查杂交果蝇的F1代，从中挑选眼色为橘黄色的果蝇，可初步认定为转基因果蝇。
(5)将转基因鉴定成功的带有橘黄色眼标记的gRNA果蝇与nos-Cas9或vasa-Cas9果蝇(能表达CAS9)杂交，方案如下(以lncRNA在2号染色体，注射86F为例)，获得非致死纯合子转基因果蝇：
5.突变体果蝇的PCR检测及测序
提取非致死纯合子转基因果蝇的DNA，设计引物，使得每对引物包含gRNA靶标位点，通过PCR扩增出目的条带，然后利用设计初时选择的限制性内切酶对上述片段进行酶切反应，检测2条染色体的序列缺失情况。如果有一条染色体发生序列突变，酶切则失败，反之则反。Secp43的扩增引物如下：
F：TACAGCTGATAGTCGTCCAC
Secp43(CG15440)
R：GACACCAGGTCAGCATTTGA
实施例2
(1)设计引物，使得每对引物包含gRNA靶标位点，通过PCR扩增该片段。由于突变位置具有酶切位点，因此仅有一条染色体的PCR片段会被顺利酶切，另外一条发生突变的染色体的PCR产物则不会被酶切。结果表明，我们获得了Secp43的杂合突变体，如图1所示。结果表明，突变体产生率约为10％，最后通过测序确定缺失位置和缺失碱基数。
(2)随机挑选羽化相同天数的野生型黑腹果蝇w1118、Secp43的移码突变体Secp43L2-6和Secp43L5-4、Secp43的非移码突变体Secp43L3-3和Secp43L10-2各30只刚羽化的成虫(雌成虫和雄成虫)，分别单头与3只w1118进行交配，3D后统计其生育能力。实验结果显示(图2)，Secp43的移码突变体雄性成虫后代数为零，而移码突变体雌性成虫的后代数与野生型之间并无显著差异。此外，Secp43的非移码突变体Secp43L3-3和Secp43L10-2的雄成虫与雌成虫的后代数与野生型相比则无明显变化，表明Secp43对雄性黑腹果蝇的生育能力起关键作用。
(3)挑选充分交配的野生型黑腹果蝇w1118、Secp43的移码突变体Secp43L2-6，解剖其精巢，观察精子的形成和发育状况，详见图3。我们发现，正常野生型果蝇的精巢内精子的分布均匀且充盈，16个初级精母细胞经过两轮减数分裂形成64个成束精子并分离，分离后的成熟精子可以进入储精囊，然而，Secp43的移码突变体中，成束精子无法完成分离，未成熟的精子则无法进入储精囊，我们推测这是雄性不育的主要原因。
上述实施例为本发明较佳的实施方式，但本发明的实施方式并不受上述实施例的限制，其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均因为等效的置换方式，都包含在本发明的保护范围之内。
序列表
<110> 广东省农业科学院植物保护研究所
<120> 蛋白Secp43或其编码基因在调控黑腹果蝇雄性生育中的应用
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1011
<212> DNA
<213> 黑腹果蝇(Drosophila melanogaster)
<400> 1
atggcgtccg tgcattgtca actgtggatg ggaagcctgg agtcctacat gacggagaac 60
ttcataatcg ccgctttccg gaagatgggc gaagatccca ccacggtgcg cctgatgcgc 120
aacaagtaca cgggcgaacc ggccggctac tgcttcgtca acttcatatc cgatgaccat 180
gcgctggacg cgatgcacaa gctaaacggt aagcccattc cgggcaccaa tcccattgtg 240
cgattccgtc ttaactcggc cagcaactcg tacaagctgc ccgggaatga acgcgaattt 300
agcgtctggg tgggcgacct cagctcggac gtggacgact atcagctgta caaggtgttc 360
tccagcaagt tcacatcgat caaaaccgcc aaggtgatct tggacagcct ggggttttcc 420
aagggttacg gctttgtgcg atttggcatc gaggatgagc agaaatcagc gctgtacgac 480
atgaacgggt acatcggatt gggtaccaag cccataaaga tttgcaatgc cgtgcccaaa 540
cccaaatccg agttgggtgg tgctgtgggc gaaggcaaca caaactatgg atatggaagc 600
ggtatgactg cagcaggtgg caccgactac tcgcagtact acgaccccac cagcacctat 660
tggcagggat accaggcctg gcaaggttac tacgagcagg ccggtgcttc gatcacagat 720
gcggctgcat actaccagca ggccatgtcg cagtcgcact ctaatccgca gactctggcc 780
cagcacgccg aggcctggag cgcacaacgc agcgcccagt acgagcagca gcagcagcag 840
cagaccgctt ccgccgccaa tggagctgag gatgagaacg gactggtgga gcacaagttc 900
gtcctggatg tggacaaact gaaccgagag gccattgatg ccgaccgccg tctgtacgat 960
gctctggaga gctctaagtg gctgcccatt gagcagctgg aagtctttta g 1011
<210> 2
<211> 336
<212> PRT
<213> 黑腹果蝇(Drosophila melanogaster)
<400> 2
Met Ala Ser Val His Cys Gln Leu Trp Met Gly Ser Leu Glu Ser Tyr
1 5 10 15
Met Thr Glu Asn Phe Ile Ile Ala Ala Phe Arg Lys Met Gly Glu Asp
20 25 30
Pro Thr Thr Val Arg Leu Met Arg Asn Lys Tyr Thr Gly Glu Pro Ala
35 40 45
Gly Tyr Cys Phe Val Asn Phe Ile Ser Asp Asp His Ala Leu Asp Ala
50 55 60
Met His Lys Leu Asn Gly Lys Pro Ile Pro Gly Thr Asn Pro Ile Val
65 70 75 80
Arg Phe Arg Leu Asn Ser Ala Ser Asn Ser Tyr Lys Leu Pro Gly Asn
85 90 95
Glu Arg Glu Phe Ser Val Trp Val Gly Asp Leu Ser Ser Asp Val Asp
100 105 110
Asp Tyr Gln Leu Tyr Lys Val Phe Ser Ser Lys Phe Thr Ser Ile Lys
115 120 125
Thr Ala Lys Val Ile Leu Asp Ser Leu Gly Phe Ser Lys Gly Tyr Gly
130 135 140
Phe Val Arg Phe Gly Ile Glu Asp Glu Gln Lys Ser Ala Leu Tyr Asp
145 150 155 160
Met Asn Gly Tyr Ile Gly Leu Gly Thr Lys Pro Ile Lys Ile Cys Asn
165 170 175
Ala Val Pro Lys Pro Lys Ser Glu Leu Gly Gly Ala Val Gly Glu Gly
180 185 190
Asn Thr Asn Tyr Gly Tyr Gly Ser Gly Met Thr Ala Ala Gly Gly Thr
195 200 205
Asp Tyr Ser Gln Tyr Tyr Asp Pro Thr Ser Thr Tyr Trp Gln Gly Tyr
210 215 220
Gln Ala Trp Gln Gly Tyr Tyr Glu Gln Ala Gly Ala Ser Ile Thr Asp
225 230 235 240
Ala Ala Ala Tyr Tyr Gln Gln Ala Met Ser Gln Ser His Ser Asn Pro
245 250 255
Gln Thr Leu Ala Gln His Ala Glu Ala Trp Ser Ala Gln Arg Ser Ala
260 265 270
Gln Tyr Glu Gln Gln Gln Gln Gln Gln Thr Ala Ser Ala Ala Asn Gly
275 280 285
Ala Glu Asp Glu Asn Gly Leu Val Glu His Lys Phe Val Leu Asp Val
290 295 300
Asp Lys Leu Asn Arg Glu Ala Ile Asp Ala Asp Arg Arg Leu Tyr Asp
305 310 315 320
Ala Leu Glu Ser Ser Lys Trp Leu Pro Ile Glu Gln Leu Glu Val Phe
325 330 335
<210> 3
<211> 19
<212> DNA
<213> 黑腹果蝇(Drosophila melanogaster)
<400> 3
atgggcgaag atcccacca 19
Claims (5)
1.蛋白Secp43或其编码基因Secp43在调控黑腹果蝇雄性生育中的应用，所述的蛋白Secp43，其氨基酸序列如SEQ ID NO.2所示。
2.根据权利要求1所述的应用，其特征在于，所述的蛋白Secp43的编码基因Secp43，其核苷酸序列如SEQ ID NO.1所示。
3.敲除编码基因Secp43在调控黑腹果蝇雄性不育中的应用，所述的编码基因Secp43，其核苷酸序列如SEQ ID NO.1所示。
4.根据权利要求3所述的应用，其特征在于，所述的敲除编码基因Secp43是利用Crispr/Cas9特异性敲除编码基因Secp43，其中用于靶向编码基因Secp43的gRNA，其核苷酸序列如SEQ ID NO.3所示。
5.gRNA在制备敲除编码基因Secp43从而调控黑腹果蝇雄性不育的制剂中的应用，所述的gRNA，其核苷酸序列如SEQ ID NO.3所示。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011353476.5A CN112480229B (zh) | 2020-11-26 | 2020-11-26 | 蛋白Secp43或其编码基因在调控黑腹果蝇雄性生育中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011353476.5A CN112480229B (zh) | 2020-11-26 | 2020-11-26 | 蛋白Secp43或其编码基因在调控黑腹果蝇雄性生育中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112480229A CN112480229A (zh) | 2021-03-12 |
| CN112480229B true CN112480229B (zh) | 2021-07-30 |
Family
ID=74935674
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011353476.5A Active CN112480229B (zh) | 2020-11-26 | 2020-11-26 | 蛋白Secp43或其编码基因在调控黑腹果蝇雄性生育中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112480229B (zh) |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1307042A (zh) * | 2000-01-28 | 2001-08-08 | 上海博道基因技术有限公司 | 一种新的多肽——人硒代半胱氨酸tRNA特异性相关蛋白SECp43,32和编码这种多肽的多核苷酸 |
| WO2011069528A1 (en) * | 2009-12-09 | 2011-06-16 | Curevac Gmbh | Lyophilization of nucleic acids in lactate-containing solutions |
-
2020
- 2020-11-26 CN CN202011353476.5A patent/CN112480229B/zh active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN112480229A (zh) | 2021-03-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US12024543B2 (en) | Micropeptides and use of same for modulating gene expression | |
| Landon et al. | Solution structure of drosomycin, the first inducible antifungal protein from insects | |
| US20180223309A1 (en) | Copi coatomer gamma subunit nucleic acid molecules that confer resistance to coleopteran and hemipteran pests | |
| US20200224214A1 (en) | Copi coatomer alpha subunit nucleic acid molecules that confer resistance to coleopteran and hemipteran pests | |
| US20200224215A1 (en) | Copi coatomer delta subunit nucleic acid molecules that confer resistance to coleopteran and hemipteran pests | |
| KR20170066404A (ko) | 딱정벌레류 및 노린재류 해충에 대한 저항성을 부여하는 copi 코토머 베타 서브유닛 핵산 분자 | |
| CN103403162A (zh) | 赋予对鞘翅目有害生物的抗性的核酸分子 | |
| CN103403164A (zh) | 靶向rho1小gtp结合蛋白并赋予对鞘翅目有害生物的抗性的核酸分子 | |
| US20150322456A1 (en) | Dre4 nucleic acid molecules that confer resistance to coleopteran pests | |
| KR20160093727A (ko) | 딱정벌레류 및/또는 노린재류 해충에 대한 저항성을 부여하는 ras 오포지트 (rop) 및 관련 핵산 분자 | |
| CN107208098A (zh) | 用于控制鞘翅目害虫染色质重塑基因亲代rnai抑制 | |
| KR20170013885A (ko) | 딱정벌레류 및 노린재류 해충에 대한 저항성을 부여하는 sec23 핵산 분자 | |
| US20180305713A1 (en) | Parental rnai suppression of chromatin remodeling genes to control coleopteran pests | |
| KR20160093728A (ko) | 딱정벌레류 해충에 대한 저항성을 부여하는 rnapii-140 핵산 분자 | |
| CN110042167B (zh) | 热激蛋白基因作为分子标记物检测昆虫温度耐受性的方法 | |
| CN110452911A (zh) | 玉米ATP结合盒转运体蛋白E2基因ZmABCE2及应用 | |
| CN112480229B (zh) | 蛋白Secp43或其编码基因在调控黑腹果蝇雄性生育中的应用 | |
| Sowden et al. | The embryonic RNA helicase gene (ERH): a new member of the DEAD box family of RNA helicases | |
| CN110747199B (zh) | 蜜蜂抗逆相关基因nf-y及其应用 | |
| CN112029777B (zh) | 一种降低水稻结实率的OsALIS4基因及其编码得到的蛋白和应用 | |
| CN110106175B (zh) | 一种dsRNA及其在害虫防治中的应用 | |
| CN112175055A (zh) | 分离的蛋白、核酸及其应用 | |
| CN105420244B (zh) | 周氏啮小蜂热激蛋白Hsp90的cDNA全长序列及应用 | |
| CN116003563B (zh) | 钙调素结合蛋白CaMBP13在调控植物耐冷性中的应用 | |
| CN110241130A (zh) | 控制植物粒数和粒重的gsn1基因、编码蛋白及其应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |