Evolutionary Rate Covariation Identifies New Members of a Protein Network Required for Female Post-Mating Responses


Seminal fluid proteins transferred from males to females during copulation are required for full fertility and can exert dramatic effects on female physiology and behavior. In Drosophila melanogaster, the seminal protein sex peptide (SP) affects mated females by increasing egg production and decreasing receptivity to courtship. These behavioral changes persist for several days because SP binds to sperm that are stored in the female. SP is then gradually released, allowing it to interact with its female-expressed receptor. The binding of SP to sperm requires five additional seminal proteins, which act together in a network. Hundreds of uncharacterized male and female proteins have been identified in this species, but individually screening each protein for network function would present a logistical challenge. To prioritize the screening of these proteins for involvement in the SP network, we used a comparative genomic method to identify candidate proteins whose evolutionary rates across the Drosophila phylogeny co-vary with those of the SP network proteins. Subsequent functional testing of 18 co-varying candidates by RNA interference identified three male seminal proteins and three female reproductive tract proteins that are each required for the long-term persistence of SP responses in females. Molecular genetic analysis showed the three new male proteins are required for the transfer of other network proteins to females and for SP to become bound to sperm that are stored in mated females. The three female proteins, in contrast, act downstream of SP binding and sperm storage. These findings expand the number of seminal proteins required for SP's actions in the female and show that multiple female proteins are necessary for the SP response. Furthermore, our functional analyses demonstrate that evolutionary rate covariation is a valuable predictive tool for identifying candidate members of interacting protein networks.


Vyšlo v časopise: Evolutionary Rate Covariation Identifies New Members of a Protein Network Required for Female Post-Mating Responses. PLoS Genet 10(1): e32767. doi:10.1371/journal.pgen.1004108
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1004108

Souhrn

Seminal fluid proteins transferred from males to females during copulation are required for full fertility and can exert dramatic effects on female physiology and behavior. In Drosophila melanogaster, the seminal protein sex peptide (SP) affects mated females by increasing egg production and decreasing receptivity to courtship. These behavioral changes persist for several days because SP binds to sperm that are stored in the female. SP is then gradually released, allowing it to interact with its female-expressed receptor. The binding of SP to sperm requires five additional seminal proteins, which act together in a network. Hundreds of uncharacterized male and female proteins have been identified in this species, but individually screening each protein for network function would present a logistical challenge. To prioritize the screening of these proteins for involvement in the SP network, we used a comparative genomic method to identify candidate proteins whose evolutionary rates across the Drosophila phylogeny co-vary with those of the SP network proteins. Subsequent functional testing of 18 co-varying candidates by RNA interference identified three male seminal proteins and three female reproductive tract proteins that are each required for the long-term persistence of SP responses in females. Molecular genetic analysis showed the three new male proteins are required for the transfer of other network proteins to females and for SP to become bound to sperm that are stored in mated females. The three female proteins, in contrast, act downstream of SP binding and sperm storage. These findings expand the number of seminal proteins required for SP's actions in the female and show that multiple female proteins are necessary for the SP response. Furthermore, our functional analyses demonstrate that evolutionary rate covariation is a valuable predictive tool for identifying candidate members of interacting protein networks.


Zdroje

1. PoianiA (2006) Complexity of seminal fluid: a review. Behavioral Ecology and Sociobiology 60: 289–310.

2. Avila FW, Sirot LK, LaFlamme BA, Rubinstein CD, Wolfner MF (2011) Insect seminal fluid proteins: Identification and function. In: Berenbaum MR, Carde RT, Robinson GE, editors. Annual Review of Entomology, Vol 56. pp. 21–40.

3. ChapmanT, DaviesSJ (2004) Functions and analysis of the seminal fluid proteins of male Drosophila melanogaster fruit flies. Peptides 25: 1477–1490.

4. HerndonLA, WolfnerMF (1995) A Drosophila seminal fluid protein, Acp26Aa, stimulates egg-laying in females for 1 day after mating. Proceedings of the National Academy of Sciences of the United States of America 92: 10114–10118.

5. SollerM, BownesM, KubliE (1997) Mating and sex peptide stimulate the accumulation of yolk in oocytes of Drosophila melanogaster. European Journal of Biochemistry 243: 732–738.

6. SollerM, BownesM, KubliE (1999) Control of oocyte maturation in sexually mature Drosophila females. Developmental Biology 208: 337–351.

7. AvilaFW, WolfnerMF (2009) Acp36DE is required for uterine conformational changes in mated Drosophila females. Proceedings of the National Academy of Sciences of the United States of America 106: 15796–15800.

8. Bloch QaziMC, WolfnerMF (2003) An early role for the Drosophila melanogaster male seminal protein Acp36DE in female sperm storage. Journal of Experimental Biology 206: 3521–3528.

9. NeubaumDM, WolfnerMF (1999) Mated Drosophila melanogaster females require a seminal fluid protein, Acp36DE, to store sperm efficiently. Genetics 153: 845–857.

10. WongA, AlbrightSN, GiebelJD, Ravi RamK, JiS, et al. (2008) A role for Acp29AB, a predicted seminal fluid lectin, in female sperm storage in Drosophila melanogaster. Genetics 180: 921–931.

11. PengJ, ZipperlenP, KubliE (2005) Drosophila sex peptide stimulates female innate immune system after mating via the Toll and Imd pathways. Current Biology 15: 1690–1694.

12. ShortSM, LazzaroBP (2010) Female and male genetic contributions to post-mating immune defence in female Drosophila melanogaster. Proceedings of the Royal Society B-Biological Sciences 277: 3649–3657.

13. CarvalhoGB, KapahiP, AndersonDJ, BenzerS (2006) Allocrine modulation of feeding behavior by the sex peptide of Drosophila. Current Biology 16: 692–696.

14. CognigniP, BaileyAP, Miguel-AliagaI (2011) Enteric neurons and systemic signals couple nutritional and reproductive status with intestinal homeostasis. Cell Metabolism 13: 92–104.

15. IsaacRE, LiC, LeedaleAE, ShirrasAD (2010) Drosophila male sex peptide inhibits siesta sleep and promotes locomotor activity in the post-mated female. Proceedings of the Royal Society B-Biological Sciences 277: 65–70.

16. Apger-McGlaughonJ, WolfnerMF (2013) Post-mating change in excretion by mated Drosophila melanogaster females is a long-term response that depends on sex peptide and sperm. Journal of Insect Physiology 59: 1024–30.

17. ChapmanT, BanghamJ, VintiG, SeifriedB, LungO, et al. (2003) The sex peptide of Drosophila melanogaster: Female post-mating responses analyzed by using RNA interference. Proceedings of the National Academy of Sciences of the United States of America 100: 9923–9928.

18. LiuHF, KubliE (2003) Sex-peptide is the molecular basis of the sperm effect in Drosophila melanogaster. Proceedings of the National Academy of Sciences of the United States of America 100: 9929–9933.

19. PengJ, ChenS, BusserS, LiuHF, HoneggerT, et al. (2005) Gradual release of sperm bound sex-peptide controls female postmating behavior in Drosophila. Current Biology 15: 207–213.

20. Ravi RamK, WolfnerMF (2007) Sustained post-mating response in D. melanogaster requires multiple seminal fluid proteins. Plos Genetics 3: e238.

21. Ravi RamK, WolfnerMF (2009) A network of interactions among seminal proteins underlies the long-term postmating response in Drosophila. Proceedings of the National Academy of Sciences of the United States of America 106: 15384–15389.

22. Ravi RamK, JiS, WolfnerMF (2005) Fates and targets of male accessory gland proteins in mated female Drosophila melanogaster. Insect Biochemistry and Molecular Biology 35: 1059–1071.

23. HasemeyerM, YapiciN, HeberleinU, DicksonBJ (2009) Sensory neurons in the Drosophila genital tract regulate female reproductive behavior. Neuron 61: 511–518.

24. RezavalC, PavlouHJ, DornanAJ, ChanY-B, KravitzEA, et al. (2012) Neural circuitry underlying Drosophila female postmating behavioral responses. Current Biology 22: 1155–1165.

25. YangCH, RumpfS, XiangY, GordonMD, SongW, et al. (2009) Control of the postmating behavioral switch in Drosophila females by internal sensory neurons. Neuron 61: 519–526.

26. YapiciN, KimYJ, RibeiroC, DicksonBJ (2008) A receptor that mediates the post-mating switch in Drosophila reproductive behaviour. Nature 451: 33–U31.

27. AvilaFW, RamKR, QaziMCB, WolfnerMF (2010) Sex peptide is required for the efficient release of stored sperm in mated Drosophila females. Genetics 186: 595–600.

28. LaFlammeBA, RamKR, WolfnerMF (2012) The Drosophila melanogaster seminal fluid protease “seminase” regulates proteolytic and post-mating reproductive processes. Plos Genetics 8: e1002435 doi:10.1371/journal.pgen.1002435

29. MuellerJL, LinklaterJR, Ravi RamK, ChapmanT, WolfnerMF (2008) Targeted gene deletion and phenotypic analysis of the Drosophila melanogaster seminal fluid protease inhibitor Acp62F. Genetics 178: 1605–1614.

30. DorusS, BusbySA, GerikeU, ShabanowitzJ, HuntDF, et al. (2006) Genomic and functional evolution of the Drosophila melanogaster sperm proteome. Nature Genetics 38: 1440–1445.

31. WasbroughER, DorusS, HesterS, Howard-MurkinJ, LilleyK, et al. (2010) The Drosophila melanogaster sperm proteome-II (DmSP-II). Journal of Proteomics 73: 2171–2185.

32. FindlayGD, MacCossMJ, SwansonWJ (2009) Proteomic discovery of previously unannotated, rapidly evolving seminal fluid genes in Drosophila. Genome Research 19: 886–896.

33. FindlayGD, YiX, MacCossMJ, SwansonWJ (2008) Proteomics reveals novel Drosophila seminal fluid proteins transferred at mating. Plos Biology 6: 1417–1426.

34. Ravi RamK, WolfnerMF (2007) Seminal influences: Drosophila Acps and the molecular interplay between males and females during reproduuction. Integrative and Comparative Biology 47: 427–445.

35. SwansonWJ, ClarkAG, Waldrip-DailHM, WolfnerMF, AquadroCF (2001) Evolutionary EST analysis identifies rapidly evolving male reproductive proteins in Drosophila. Proceedings of the National Academy of Sciences of the United States of America 98: 7375–7379.

36. AllenAK, SpradlingAC (2008) The Sf1-related nuclear hormone receptor Hr39 regulates Drosophila female reproductive tract development and function. Development 135: 311–321.

37. ArbeitmanMN, FlemingAA, SiegalML, NullBH, BakerBS (2004) A genomic analysis of Drosophila somatic sexual differentiation and its regulation. Development 131: 2007–2021.

38. ProkupekA, HoffmannF, EyunSI, MoriyamaE, ZhouM, et al. (2008) An evolutionary expressed sequence tag analysis of Drosophila spermatheca genes. Evolution 62: 2936–2947.

39. ProkupekAM, EyunSI, KoL, MoriyamaEN, HarshmanLG (2010) Molecular evolutionary analysis of seminal receptacle sperm storage organ genes of Drosophila melanogaster. Journal of Evolutionary Biology 23: 1386–1398.

40. ProkupekAM, KachmanSD, LadungaI, HarshmanLG (2009) Transcriptional profiling of the sperm storage organs of Drosophila melanogaster. Insect Molecular Biology 18: 465–475.

41. ClarkNL, AlaniE, AquadroCF (2012) Evolutionary rate covariation reveals shared functionality and coexpression of genes. Genome Research 22: 714–720.

42. DrummondDA, RavalA, WilkeCO (2006) A single determinant dominates the rate of yeast protein evolution. Molecular Biology and Evolution 23: 327–337.

43. LarracuenteAM, SacktonTB, GreenbergAJ, WongA, SinghND, et al. (2008) Evolution of protein-coding genes in Drosophila. Trends in Genetics 24: 114–123.

44. LiaoB-Y, ScottNM, ZhangJ (2006) Impacts of gene essentiality, expression pattern, and gene compactness on the evolutionary rate of mammalian proteins. Molecular Biology and Evolution 23: 2072–2080.

45. McInerneyJO (2006) The causes of protein evolutionary rate variation. Trends in Ecology & Evolution 21: 230–232.

46. MintserisJ, WengZP (2005) Structure, function, and evolution of transient and obligate protein-protein interactions. Proceedings of the National Academy of Sciences of the United States of America 102: 10930–10935.

47. PalC, PappB, HurstLD (2001) Highly expressed genes in yeast evolve slowly. Genetics 158: 927–931.

48. PalC, PappB, LercherMJ (2006) An integrated view of protein evolution. Nature Reviews Genetics 7: 337–348.

49. RochaEPC, DanchinA (2004) An analysis of determinants of amino acids substitution rates in bacterial proteins. Molecular Biology and Evolution 21: 108–116.

50. ClarkNL, GasperJ, SekinoM, SpringerSA, AquadroCF, et al. (2009) Coevolution of interacting fertilization proteins. Plos Genetics 5: e1000570.

51. HakesL, LovellSC, OliverSG, RobertsonDL (2007) Specificity in protein interactions and its relationship with sequence diversity and coevolution. Proceedings of the National Academy of Sciences of the United States of America 104: 7999–8004.

52. KannMG, ShoemakerBA, PanchenkoAR, PrzytyckaTM (2009) Correlated evolution of interacting proteins: Looking behind the mirrortree. Journal of Molecular Biology 385: 91–98.

53. LovellSC, RobertsonDL (2010) An integrated view of molecular coevolution in protein-protein interactions. Molecular Biology and Evolution 27: 2567–2575.

54. ClarkNL, AlaniE, AquadroCF (2013) Evolutionary rate covariation in meiotic proteins results from fluctuating evolutionary pressure in yeasts and mammals. Genetics 193: 529–538.

55. ClarkNL, AquadroCF (2010) A novel method to detect proteins evolving at correlated rates: identifying new functional relationships between coevolving proteins. Molecular Biology and Evolution 27: 1152–1161.

56. GohCS, CohenFE (2002) Co-evolutionary analysis reveals insights into protein-protein interactions. Journal of Molecular Biology 324: 177–192.

57. PazosF, ValenciaA (2001) Similarity of phylogenetic trees as indicator of protein-protein interaction. Protein Engineering 14: 609–614.

58. SatoT, YamanishiY, KanehisaM, TohH (2005) The inference of protein-protein interactions by co-evolutionary analysis is improved by excluding the information about the phylogenetic relationships. Bioinformatics 21: 3482–3489.

59. JuanD, PazosF, ValenciaA (2008) High-confidence prediction of global interactomes based on genome-wide coevolutionary networks. Proceedings of the National Academy of Sciences of the United States of America 105: 934–939.

60. TabachY, BilliAC, HayesGD, NewmanMA, ZukO, et al. (2013) Identification of small RNA pathway genes using patterns of phylogenetic conservation and divergence. Nature 493: 694–698.

61. ConsortiumDG (2007) Evolution of genes and genomes on the Drosophila phylogeny. Nature 450: 203–218.

62. KimYJ, BartalskaK, AudsleyN, YamanakaN, YapiciN, et al. (2010) MIPs are ancestral ligands for the sex peptide receptor. Proceedings of the National Academy of Sciences of the United States of America 107: 6520–6525.

63. PoelsJ, Van LoyT, VandersmissenHP, Van HielB, Van SoestS, et al. (2010) Myoinhibiting peptides are the ancestral ligands of the promiscuous Drosophila sex peptide receptor. Cellular and Molecular Life Sciences 67: 3511–3522.

64. ChintapalliVR, WangJ, DowJAT (2007) Using FlyAtlas to identify better Drosophila melanogaster models of human disease. Nature Genetics 39: 715–720.

65. ArnoneJT, Robbins-PiankaA, AraceJR, Kass-GergiS, McAlearMA (2012) The adjacent positioning of co-regulated gene pairs is widely conserved across eukaryotes. BMC Genomics 13: 546 doi:10.1186/1471-2164-13-546

66. HuY, RoeselC, FlockhartI, PerkinsL, PerrimonN, et al. (2013) UP-TORR: Online tool for accurate and up-to-date annotation of RNAi reagents. Genetics 195: 37–45.

67. DeanMD, ClarkNL, FindlayGD, KarnRC, YiXH, et al. (2009) Proteomics and comparative genomic investigations reveal heterogeneity in evolutionary rate of male reproductive proteins in mice (Mus domesticus). Molecular Biology and Evolution 26: 1733–1743.

68. PilchB, MannM (2005) Large scale proteomic analysis of human seminal plasma. Molecular & Cellular Proteomics 4: S205–S205.

69. SirotLK, HardstoneMC, HelinskiMEH, RibeiroJMC, KimuraM, et al. (2011) Towards a semen proteome of the dengue vector mosquito: Protein identification and potential functions. Plos Neglected Tropical Diseases 5: e989 doi:10.1371/journal.pntd.0000989

70. RossJ, JiangH, KanostMR, WangY (2003) Serine proteases and their homologs in the Drosophila melanogaster genome: an initial analysis of sequence conservation and phylogenetic relationships. Gene 304: 117–131.

71. CarpentierM, GuillemetteC, BaileyJL, BoileauG, JeannotteL, et al. (2004) Reduced fertility in male mice deficient in the zinc metallopeptidase NL1. Molecular and Cellular Biology 24: 4428–4437.

72. PintadoCO, PintoFM, PennefatherJN, HidalgoA, BaamondeA, et al. (2003) A role for tachykinins in female mouse and rat reproductive function. Biology of Reproduction 69: 940–946.

73. PintoFM, ArmestoCP, MagranerJ, TrujilloM, MartinJD, et al. (1999) Tachykinin receptor and neutral endopeptidase gene expression in the rat uterus: Characterization and regulation in response to ovarian steroid treatment. Endocrinology 140: 2526–2532.

74. PetersenTN, BrunakS, von HeijneG, NielsenH (2011) SignalP 4.0: discriminating signal peptides from transmembrane regions. Nature Methods 8: 785–786.

75. Marchler-BauerA, LuS, AndersonJB, ChitsazF, DerbyshireMK, et al. (2011) CDD: a Conserved Domain Database for the functional annotation of proteins. Nucleic Acids Research 39: D225–D229.

76. KelleyLA, SternbergMJE (2009) Protein structure prediction on the Web: a case study using the Phyre server. Nature Protocols 4: 363–371.

77. RocheN, KaufmanT (1996) Characterization of the homoeotic target gene Epidermal stripes and patches, a Drosophila homologue of the human diastrophic dysplasia gene. Molecular Biology of the Cell 7: 699–699.

78. BassettAR, TibbitC, PontingCP, LiuJ-L (2013) Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system. Cell Reports 4: 220–228.

79. GratzSJ, CummingsAM, NguyenJN, HammDC, DonohueLK, et al. (2013) Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease. Genetics 194: 1029–1035.

80. ChanHC, RuanYC, HeQ, ChenMH, ChenH, et al. (2009) The cystic fibrosis transmembrane conductance regulator in reproductive health and disease. Journal of Physiology 587: 2187–2195.

81. LaFlammeBA, WolfnerMF (2013) Identification and function of proteolysis regulators in seminal fluid. Molecular Reproduction and Development 80: 80–101.

82. Ravi RamK, SirotLK, WolfnerMF (2006) Predicted seminal astacin-like protease is required for processing of reproductive proteins in Drosophila melanogaster. Proceedings of the National Academy of Sciences of the United States of America 103: 18674–18679.

83. SmithJR, StanfieldGM (2011) TRY-5 Is a sperm-activating protease in Caenorhabditis elegans aeminal fluid. Plos Genetics 7: e1002375 doi:10.1371/journal.pgen.1002375

84. StanfieldGM, VilleneuveAM (2006) Regulation of sperm activation by SWM-1 is required for reproductive success of C. elegans males. Current Biology 16: 252–263.

85. ZhaoY, SunW, ZhangP, ChiH, ZhangM-J, et al. (2012) Nematode sperm maturation triggered by protease involves sperm-secreted serine protease inhibitor (Serpin). Proceedings of the National Academy of Sciences of the United States of America 109: 1542–1547.

86. LungO, WolfnerMF (2001) Identification and characterization of the major Drosophila melanogaster mating plug protein. Insect Biochemistry and Molecular Biology 31: 543–551.

87. GligorovD, SitnikJL, MaedaRK, WolfnerMF, KarchF (2013) A Novel Function for the Hox Gene Abd-B in the Male Accessory Gland Regulates the Long-Term Female Post-Mating Response in Drosophila. Plos Genetics 9: e1003395 doi:10.1371/journal.pgen.1003395

88. LeiblichA, MarsdenL, GandyC, CorriganL, JenkinsR, et al. (2012) Bone morphogenetic protein- and mating-dependent secretory cell growth and migration in the Drosophila accessory gland. Proceedings of the National Academy of Sciences of the United States of America 109: 19292–19297.

89. ChenPS, Stumm-ZollingerE, AigakiT, BalmerJ, BienzM, et al. (1988) A male accessory gland peptide that regulates reproductive behavior of female Drosophila melanogaster. Cell 54: 291–298.

90. HaninO, AzrielliA, ZakinV, ApplebaumS, RafaeliA (2011) Identification and differential expression of a sex-peptide receptor in Helicoverpa armigera. Insect Biochemistry and Molecular Biology 41: 537–544.

91. DottoriniT, NicolaidesL, RansonH, RogersDW, CrisantiA, et al. (2007) A genome-wide analysis in Anopheles gambiae mosquitoes reveals 46 male accessory gland genes, possible modulators of female behavior. Proceedings of the National Academy of Sciences of the United States of America 104: 16215–16220.

92. MarkowTA (1996) Evolution of Drosophila mating systems. Evolutionary Biology, Vol 29 29: 73–106.

93. ThailayilJ, MagnussonK, GodfrayHCJ, CrisantiA, CatterucciaF (2011) Spermless males elicit large-scale female responses to mating in the malaria mosquito Anopheles gambiae. Proceedings of the National Academy of Sciences of the United States of America 108: 13677–13681.

94. MarkowTA, O'GradyPM (2005) Evolutionary genetics of reproductive behavior in Drosophila: connecting the dots. Annual Review of Genetics 39: 263–291.

95. ClarkNL, AagaardJE, SwansonWJ (2006) Evolution of reproductive proteins from animals and plants. Reproduction 131: 11–22.

96. FindlayGD, SwansonWJ (2010) Proteomics enhances evolutionary and functional analysis of reproductive proteins. BioEssays 32: 26–36.

97. TurnerLM, HoekstraHE (2008) Causes and consequences of the evolution of reproductive proteins. International Journal of Developmental Biology 52: 769–780.

98. KuijperB, StewartAD, RiceWR (2006) The cost of mating rises nonlinearly with copulation frequency in a laboratory population of Drosophila melanogaster. Journal of Evolutionary Biology 19: 1795–1802.

99. RiceWR (1996) Sexually antagonistic male adaptation triggered by experimental arrest of female evolution. Nature 381: 232–234.

100. EdwardDA, FrickeC, GerrardDT, ChapmanT (2011) Quantifying the life-history response to increased male exposure in female Drosophila melanogaster. Evolution 65: 564–573.

101. WigbyS, ChapmanT (2005) Sex peptide causes mating costs in female Drosophila melanogaster. Current Biology 15: 316–321.

102. CireraS, AguadeM (1997) Evolutionary history of the sex peptide (Acp70A) gene region in Drosophila melanogaster. Genetics 147: 189–197.

103. WongA, TurchinMC, WolfnerMF, AquadroCF (2008) Evidence for positive selection on Drosophila melanogaster seminal fluid protease homologs. Molecular Biology and Evolution 25: 497–506.

104. OstlundG, SchmittT, ForslundK, KostlerT, MessinaDN, et al. (2010) InParanoid 7: new algorithms and tools for eukaryotic orthology analysis. Nucleic Acids Research 38: D196–D203.

105. EdgarRC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research 32: 1792–1797.

106. YangZ (2007) PAML 4: Phylogenetic analysis by maximum likelihood. Molecular Biology and Evolution 24: 1586–1591.

107. DietzlG, ChenD, SchnorrerF, SuKC, BarinovaY, et al. (2007) A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila. Nature 448: 151–U151.

108. NiJ-Q, ZhouR, CzechB, LiuL-P, HolderbaumL, et al. (2011) A genome-scale shRNA resource for transgenic RNAi in Drosophila. Nature Methods 8: 405–U446.

109. NiJ-Q, LiuL-P, BinariR, HardyR, ShimH-S, et al. (2009) A Drosophila resource of transgenic RNAi lines for neurogenetics. Genetics 182: 1089–1100.

110. NiJ-Q, MarksteinM, BinariR, PfeifferB, LiuL-P, et al. (2008) Vector and parameters for targeted transgenic RNA interference in Drosophila melanogaster. Nature Methods 5: 49–51.

111. XueL, NollM (2002) Dual role of the Pax gene paired in accessory gland development of Drosophila. Development 129: 339–346.

112. SchnakenbergSL, MatiasWR, SiegalML (2011) Sperm-storage defects and live birth in Drosophila females lacking spermathecal secretory cells. Plos Biology 9: e1001192 doi:10.1371/journal.pbio.1001192

113. BoutrosM, KigerAA, ArmknechtS, KerrK, HildM, et al. (2004) Genome-wide RNAi analysis of growth and viability in Drosophila cells. Science 303: 832–835.

Štítky
Genetika Reprodukčná medicína

Článok vyšiel v časopise

PLOS Genetics


2014 Číslo 1
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa