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Issue:ISSN 2095-1353
           CN 11-6020/Q
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Your Position :Home->Past Journals Catalog->2018年55 No.2

A non-destructive method of genotyping individual insects from the exuviate of final instar larvae, or puparia
Author of the article:ZHENG Mei-Yan** ZHU Guan-Heng CHEN Yan HAN Qi YANG Yi-Hua DONG Shuang-Lin***
Author's Workplace:Department of Entomology, Nanjing Agricultural University, Nanjing 210095, China
Key Words: larval exuviate, puparium, genomic DNA, PCR, mutant strain
Abstract:

[Objectives]  The single pair mating strategy is often used to screen homozygous insect strains for the functional study of insect genes. This requires genotyping individual insects without killing, or otherwise harming them, before pair combinations can be made. The purpose of this study was to evaluate a non-destructive method of genotyping individual insects using the exuviate of final instar larvae, or puparia. [Methods]  Genomic DNA was extracted from exuviates of the final instar larvae and puparia of Spodoptera litura Fabricius, Athetis lepigone Möschler and Plutella xyllostella Linnaeus with DNAiso reagent. The general odorant binding protein 1 (GOBP1) gene was used as a representative gene and amplified by PCR, followed by agarose gel electrophoresis, ligation, transformation, and TA clone sequencing. [Results]  Agarose gel electrophoresis of GOBP1 PCR products from S. litura or A. lepigone produced a bright, single band of the expected size, the identity of which was confirmed by TA cloning and sequencing. However, due to the low quantity of larval exuviate and small size of puparia, the concentration of DNA from P. xyllostella was very low and the expected target band of GOBP1 was not detected by gel electrophoresis. [Conclusion]  Insects of the same size, or larger, than S. litura and A. lepigone can be non-destructively genotyped by PCR using final instar larvae exuviate, or puparia.

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