Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA

被引:214
作者
Amdam, GV
Simoes, ZLP
Guidugli, KR
Norberg, K
Omholt, SW [1 ]
机构
[1] Agr Univ Norway, Ctr Integrat Genet, N-1432 As, Norway
[2] Agr Univ Norway, Dept Anim Sci, N-1432 As, Norway
[3] Univ Sao Paulo, Dept Biol, BR-14040901 Ribeirao Preto, Brazil
[4] Univ Sao Paulo, Dept Genet, BR-14040901 Ribeirao Preto, Brazil
关键词
D O I
10.1186/1472-6750-3-1
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Background: The ability to manipulate the genetic networks underlying the physiological and behavioural repertoires of the adult honeybee worker (Apis mellifera) is likely to deepen our understanding of issues such as learning and memory generation, ageing, and the regulatory anatomy of social systems in proximate as well as evolutionary terms. Here we assess two methods for probing gene function by RNA interference (RNAi) in adult honeybees. Results: The vitellogenin gene was chosen as target because its expression is unlikely to have a phenotypic effect until the adult stage in bees. This allowed us to introduce dsRNA in preblastoderm eggs without affecting gene function during development. Of workers reared from eggs injected with dsRNA derived from a 504 bp stretch of the vitellogenin coding sequence, 15% had strongly reduced levels of vitellogenin mRNA. When dsRNA was introduced by intra-abdominal injection in newly emerged bees, almost all individuals ( 96%) showed the mutant phenotype. An RNA-fragment with an apparent size similar to the template dsRNA was still present in this group after 15 days. Conclusion: Injection of dsRNA in eggs at the preblastoderm stage seems to allow disruption of gene function in all developmental stages. To dissect gene function in the adult stage, the intra-abdominal injection technique seems superior to egg injection as it gives a much higher penetrance, it is much simpler, and it makes it possible to address genes that are also expressed in the embryonic, larval or pupal stages.
引用
收藏
页数:8
相关论文
共 40 条
  • [1] The regulatory anatomy of honeybee lifespan
    Amdam, GV
    Omholt, SW
    [J]. JOURNAL OF THEORETICAL BIOLOGY, 2002, 216 (02) : 209 - 228
  • [2] AMDAM GV, P NATL ACAD SCI US
  • [3] Apolipophorin II/I, apolipoprotein B, vitellogenin, and microsomal triglyceride transfer protein genes are derived from a common ancestor
    Babin, PJ
    Bogerd, J
    Kooiman, FP
    Van Marrewijk, WJA
    Van der Horst, DJ
    [J]. JOURNAL OF MOLECULAR EVOLUTION, 1999, 49 (01) : 150 - 160
  • [4] Barchuk Angel Roberto, 2002, Journal of Insect Science (Tucson), V2, P1
  • [5] Role for a bidentate ribonuclease in the initiation step of RNA interference
    Bernstein, E
    Caudy, AA
    Hammond, SM
    Hannon, GJ
    [J]. NATURE, 2001, 409 (6818) : 363 - 366
  • [6] A gridded genomic library of the honeybee (Apis mellifera):: A reference library system for basic and comparative genetic studies of a hymenopteran genome
    Beye, M
    Poch, A
    Burgtorf, C
    Moritz, RFA
    Lehrach, H
    [J]. GENOMICS, 1998, 49 (02) : 317 - 320
  • [7] Specific developmental gene silencing in the honey bee using a homeobox motif
    Beye, M
    Härtel, S
    Hagen, A
    Hasselmann, M
    Omholt, SW
    [J]. INSECT MOLECULAR BIOLOGY, 2002, 11 (06) : 527 - 532
  • [8] The relationship between level of pollen in the diet, vitellogenin and juvenile hormone titres in Africanized Apis mellifera workers
    Bitondi, MMG
    Simoes, ZLP
    [J]. JOURNAL OF APICULTURAL RESEARCH, 1996, 35 (01) : 27 - 36
  • [9] Induction of RNA interference in Caenorhabditis elegans by RNAs derived from plants exhibiting post-transcriptional gene silencing
    Boutla, A
    Kalantidis, K
    Tavernarakis, N
    Tsagris, M
    Tabler, M
    [J]. NUCLEIC ACIDS RESEARCH, 2002, 30 (07) : 1688 - 1694
  • [10] Drosophila antimicrobial peptides
    Bulet, P
    [J]. M S-MEDECINE SCIENCES, 1999, 15 (01): : 23 - 29