Method for improving egg laying amount of original strain of mulberry silkworm

A technology of silkworm and original seed, which is applied in the field of genetic engineering, can solve the problems of increasing egg production of silkworm by expressing endogenous genes, and achieve the effects of avoiding potential harm, improving egg production level, and saving production costs

Inactive Publication Date: 2013-08-14
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no report on increasing the egg production of silkworm by endogenous gene expression

Method used

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  • Method for improving egg laying amount of original strain of mulberry silkworm
  • Method for improving egg laying amount of original strain of mulberry silkworm
  • Method for improving egg laying amount of original strain of mulberry silkworm

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment one: silkworm vasa - like Gene promoter control GAL 4. UAS control Bmovo- 1 Construction of transgenic binary system

[0022] 1. Bmovo- 1 Cloning of gene cDNA

[0023] Bombyx mori larvae ovaries were dissected, total RNA was extracted, and total RNA was used as a template, and oligo(dT) was used as a primer to synthesize cDNA by reverse transcription, using SEQ ID No.1 ( gat atc atg ccg aaa atc ​​ttc tgg att aag, underlined Eco RV site) and SEQ ID No.2 ( gtc gac tta att gtg tac tgg cat ggg c, underlined Sal Ⅰ site) as primers, cDNA was amplified by conventional PCR, and a specific product of 2.42 kb was recovered; the product was cloned into pMD TM 19-T (Dalian Bao Biological Co., Ltd.), obtained from pMD-Bmovo, sequenced, attached figure 1 For the sequencing results, it can be confirmed that it is consistent with the published sequence of GenBank (accession number: GU477588).

[0024] 2. Construction of effector transgene vector pigU...

Embodiment 2

[0039] Embodiment two: silkworm nanos - like gene mover control GAL 4. UAS control Bmovo- 1 Construction of transgenic binary system

[0040] 1. Construction of activating transgenic vector pignanos-Gal4

[0041] (1) Construction of the pignanos plasmid: using the silkworm genome as a template, SEQ ID No.13 (tat ccc ggg tgc ctt aca tca aag aca ta, underlined as Sma Ⅰ site) and SEQ ID No.14 (cgc aga tct ttc aat tac gaa gat gtt gct underlined as Bgl Ⅱ site) as primers, PCR amplification, recovery of 0.6 kb specific fragments, cloned into pMD TM 19-T, obtained pMD-nanos, confirmed by sequencing and published on GenBank nanos - like Gene promoter sequence (8401-9001 region of BABH01013628.1) is consistent, pMD-nanos plasmid Sma I / Bgl Ⅱ Double enzyme digestion, recovery of the promoter fragment, cloned into the same digestion piggyA3GFP vector to obtain pignanos plasmid;

[0042] (2) Construction of pignanos-Gal4 vector: pBGT1 (refer to Lukacsovich T ...

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Abstract

The invention belongs to the field of genetic engineering, and in particular relates to a method for improving egg laying amount of an original strain of mulberry silkworm. The method comprises the following steps: by using a piggyBAC transposons carrier containing a fluorescent protein reporter gene as a basic carrier, constructing a transgenetic mulberry silkworm vlg-GAL4 system or a nanos-GAL4 system which controls GAL4 genetic expression through a vasa-like genetic promoter or a nanos-like genetic promoter through operation of genetic engineering; constructing a transgenetic mulberry silkworm UAS-ovo-1 system which controls genetic expression of mulberry silkworm ovo-1 through a UAS promoter through operation of genetic engineering; hybridizing the vlg-GAL4 system or nanos-GAL4 system with the UAS-ovo-1 system to obtain first-filial generation mulberry silkworm; and selfing and laying by the mulberry silkworm obtained, so that the egg laying amount of original strain of mulberry silkworm is improved. The method provided by the invention over-expresses mulberry silkworm ovo-1 genes in the genital gland through the GAL4/UAS binary system, so that the laying level of a single moth is improved, the laying amount is increased by over 10%, and meanwhile, the production cost of cocoons is lowered. The method has important application value.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a method for increasing the egg production of silkworm original species. Background technique [0002] Silkworm silk protein efficient synthesis, egg production, development and metamorphosis, immunity and resistance, and sex determination are the five most important traits of silkworm. Improving the egg production of silkworm original species is an important content of silkworm egg production, which has a significant impact on reducing the production cost of seedlings and improving labor productivity. The number of eggs produced by a single moth and the number of eggs laid are inherent traits of silkworm species. In production, the number of eggs produced and laid by a single moth of the silkworm can be increased to a certain extent by strengthening feeding management and being fed with good mulberry, but this conventional method is difficult to significantly incr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N15/11A01K67/04
Inventor 贡成良曹广力薛仁宇潘中华郑小坚
Owner SUZHOU UNIV
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