RNAi KNOCKDOWN OF BmRab3 LED TO LARVA AND PUPA LETHALITY IN SILKWORM Bombyx mori L.
Rab3 GTPases are known to play key a role in vesicular trafficking, and express highest in brain and endocrine tissues. In mammals, Rab3 GTPases are paralogs unlike in insect. In this study, we cloned Rab3 from the silk gland tissue of silkworm Bombyx mori, and identified it as BmRab3. Our in silico analysis indicated that BmRab3 is an isoform with a theoretical isoelectric point and molecular weight of 5.52 and 24.3 kDa, respectively. Further, BmRab3 showed the C-terminal hypervariability for GGT2 site but having two other putative guanine nucleotide exchange factor/GDP dissociation inhibitor interaction sites. Multiple alignment sequence indicated high similarities of BmRab3 with Rab3 isoforms of other species. The phylogeny tree showed BmRab3 clustered between the species of Tribolium castaneum and Aedes aegypti. Meanwhile, the expression analysis of BmRab3 showed the highest expression in middle silk glands (MSGs) than all other tissues in the third day of fifth-instar larva. Simultaneously, we showed the differential expression of BmRab3 in the early instar larva development, followed by higher expression in male than female pupae. In vivo dsRNA interference of BmRab3 reduced the expression of BmRab3 by 75% compared to the control in the MSGs in the first day. But as the worm grew to the third day, the difference of BmRab3 between knockdown and control was only about 10%. The knockdown later witnessed underdevelopment of the larvae and pharate pupae lethality in the overall development of silkworm B. mori L.
Institute of Sericulture and Apiculture, College of Animal Sciences, Zhejiang University, Hangzhou, P. R. China.
Chabungbam Orville
CO
Singh
Institute of Sericulture and Apiculture, College of Animal Sciences, Zhejiang University, Hangzhou, P. R. China.
Hu-hu
HH
Xin
Institute of Sericulture and Apiculture, College of Animal Sciences, Zhejiang University, Hangzhou, P. R. China.
Rui-ting
RT
Chen
Institute of Sericulture and Apiculture, College of Animal Sciences, Zhejiang University, Hangzhou, P. R. China.
Mei-xian
MX
Wang
Institute of Sericulture and Apiculture, College of Animal Sciences, Zhejiang University, Hangzhou, P. R. China.
Shuang
S
Liang
Institute of Sericulture and Apiculture, College of Animal Sciences, Zhejiang University, Hangzhou, P. R. China.
Yan
Y
Lu
Institute of Sericulture and Apiculture, College of Animal Sciences, Zhejiang University, Hangzhou, P. R. China.
Zi-zheng
ZZ
Cai
Institute of Sericulture and Apiculture, College of Animal Sciences, Zhejiang University, Hangzhou, P. R. China.
Deng-pan
DP
Zhang
Institute of Sericulture and Apiculture, College of Animal Sciences, Zhejiang University, Hangzhou, P. R. China.
Yun-gen
YG
Miao
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000502:physiology*; D005260:Female; D055785:Gene Knockdown Techniques; D019476:Insect Proteins / Q000502:physiology*; D007814:Larva / Q000502:physiology; D008297:Male; D008969:Molecular Sequence Data; D011679:Pupa / Q000378:metabolism; D034622:RNA Interference; D017422:Sequence Analysis, DNA; D020692:rab3 GTP-Binding Proteins / Q000502:physiology*
D019476:Insect Proteins; D020692:rab3 GTP-Binding Proteins
BmRab3; dsRNA interference; middle silk glands (MSG); silkworm Bombyx mori (B. mori)