Method for controlling domestic silkworm pupal stage growth

A silkworm and pupa stage technology, applied in the field of genetic engineering, to achieve the effects of increasing flexibility, prolonging the pupa stage, and prolonging the pupa stage

Inactive Publication Date: 2011-11-23
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, how to ensure BmKIT 3 R The specific expression of the gene in the pupal

Method used

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  • Method for controlling domestic silkworm pupal stage growth
  • Method for controlling domestic silkworm pupal stage growth
  • Method for controlling domestic silkworm pupal stage growth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment one: Bombyx mori cocoon protease gene promoter ( PDPs )control GAL 4. UAS control BmKIT 3 R Construction of transgenic binary system

[0028] 1. BmKIT3 R artificial synthesis of genes

[0029] According to the published scorpion toxin BmKIT 3 The sequence of the gene, referring to the codon preference of the silkworm, synthesized a 189 nt sequence including the start codon ATG and the stop codon TGA BmKIT 3 R The gene, specific nucleotide sequence and corresponding amino acid sequence are shown in SEQ ID No.1 and SEQ ID No.2 respectively. The synthesized sequence was cloned into pUCm-T (product of Shanghai Shenergy Biotechnology Co., Ltd.), and the recombinant plasmid pUCm-IT3R was obtained, which was confirmed to be completely consistent with the designed sequence by sequencing.

[0030] 2. Construction of effector transgene vector piggyA3GFP-ie-neo-UAS-IT3R-polyA

[0031] (1) pSK-UAS plasmid construction: using pUAST plasmid DNA (see Bra...

Embodiment 2

[0052] Example 2: Bombyx mori cocoon protease gene promoter (PDP) controls GAL4, UAS control egt Construction of transgenic binary system

[0053] The technical operation process is as follows:

[0054] 1. Construction of the activated transgenic vector pigA3GFP-ie-neo-PDP-Gal4-PA: same as Step 3 in Example 1.

[0055] 2. Construction and identification of PDP-GAL4 system: same as Step 5 of Example 1.

[0056] 3. Construction of effector transgenic vector pigA3GFP-ie-neo-UAS-EGT

[0057] (1) Construction of pSK-UAS-egt vector: Bombyx mori baculovirus (BmNPV) genomic DNA was extracted according to conventional methods. PCR upstream and downstream primers Egt-1 (SEQ ID No.17: cg c tcg ag a tga cta ttc ttt gct ggc, underlined xho Ⅰ site) and Egt-2b (SEQ ID No.18: cg g gta cc t tgg ttg cta ctg gca cat aag cgc, underlined Kpn Ⅰ site); using the BmNPV genome as a template, PCR amplification was carried out, and the 1.7 kb egt Gene fragment. Kpn I / xho After 1 double ...

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Abstract

The invention belongs to the field of genetic engineering, and specifically relates to a method of domestic silkworm transgene for controlling domestic silkworm pupal stage growth through UAS/GAL4 dual system. According to the method, piggyBAC transposon vector pigA3GFP is adopted as a base vector; through genetic engineering operation, GAL4 gene expressed transgenic domestic silkworm PDP-GAL4 system controlled by domestic silkworm cocoon protease gene promoter (PDP) is established; through genetic engineering operation, domestic silkworm baculovirus egt gene or charybdotoxin BmKIT3R gene expressed transgenic domestic silkworm UAS-egt or UAS-BmKIT3R system controlled by UAS promoter is established; the PDP-GAL4 system and the UAS-egt or UAS-BmKIT3R system are hybridized, such that larval stage growth of an F1 generation is normal, and pupal stage of the F1 generation is prolonged for 7 to 10 days. According to the invention, with an UAS/GAL4 dual system, the F1 generation silkworm pupal stage is prolonged by using domestic silkworm baculovirus egt gene or BmKIT3R gene. Flexibilities of silkworm cocoon purchasing and filature technology are improved. Therefore, the invention 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 controlling the pupal stage development of the silkworm through the UAS / GAL4 binary system of the transgenic silkworm. Background technique [0002] Efficient silk protein synthesis, development and metamorphosis, immunity and resistance, and sex determination are the four most important traits of silkworm. The artificial regulation of silkworm metamorphosis and development is an important content of silk production, and the artificial regulation of silkworm metamorphosis and development has a significant impact on the production structure and overall production efficiency of the silk industry. The silkworm is a complete metamorphosis insect, and the pupal period is very short, only 2 weeks. Since the moth mouth cocoon is not suitable for silk reeling, it is necessary to complete the acquisition and drying of fresh cocoons before the pupa turns into a m...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N15/12C12N15/866A01K67/04
Inventor 贡成良曹广力薛仁宇郑小坚潘中华
Owner SUZHOU UNIV
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