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Construction method of cultivated silkworm with controllable upgrowth and upgrowth control method

A construction method and technology of silkworm, applied in the field of preparation of transgenic silkworm

Inactive Publication Date: 2009-04-29
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above methods all interfere with the developmental metamorphosis of silkworms by applying certain chemical substances, and do not involve changing the developmental metamorphosis of silkworms through genetic manipulation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Preparation of transgenic silkworm expressing egt gene driven by the hsp20.4 promoter of silkworm heat shock protein gene

[0021] The technical operation process is as follows:

[0022] 1. Preparation of egt gene of silkworm baculovirus

[0023] Bombyx mori baculovirus (BmNPV) genomic DNA was extracted according to conventional methods. PCR upstream and downstream primers were designed according to the published complete genome sequence of BmNPV-T3 strain (GenBank accession number: L33180), primer 1: 5′-cg ctcgag atgactattctttgctggc-3' (Xho I restriction site is underlined), primer 2: 5'-cg ggtacc gaccggcacggcaagctggcgc-3' (the underline is the Kpn I restriction site). Using the BmNPV genome as a template, PCR amplification was performed under the following conditions: pre-denaturation at 95°C for 3 min, 50 sec at 95°C, 50 sec at 50°C, 90 sec at 72°C, and 10 min at 72°C after 35 cycles. The PCR amplified product was recovered with a 1% agarose gel cutti...

Embodiment 2

[0041] Embodiment 2: Larval development control of transgenic silkworm

[0042] The transgenic silkworm obtained in Example 1 was normally reared until the 5th day of the fifth instar, heat-induced at 42°C for 1 hour, and then reared normally. Compared with the non-transgenic silkworm treated in the same way, the elapsed time of the fifth instar of the transgenic silkworm was extended by 1 to 2 sky.

Embodiment 3

[0043] Example 3: Control of Pupal Stage Development of Transgenic Bombyx mori

[0044] The transgenic silkworm obtained in Example 1 was normally fed until cocooning, and on the 4th day of pupation, it was heat-induced at 42°C for 1 hour, and then protected normally. Compared with the non-transgenic silkworm chrysalis treated in the same way, the pupal age of the transgenic silkworm was prolonged by 3 days above.

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Abstract

The invention discloses a method for constructing a silkworm with controllable growth and a method for controlling growth. A transposon piggyBAC factor is used as a basic vector; a recombining transgenic vector with an egt gene expression box controlled by a silkworm heat shock protein gene promoter is constructed through gene engineering operation; the recombining transgenic vector and an auxiliary plasmid are mixed and are conducted to a first spawn; newly-hatched silkworms after incubation are subjected to routine screening to obtain silkworms with fluorescence protein report genes; the egt gene is detected and is breed to prepare a transgenic silkworm; and the breed transgenic silkworm is subjected to thermal induction at a temperature of between 41 and 43 DEG C for 0.5 to 2 hours so as to control the growth of the silkworm. The method realizes that the rhabdovirus egt gene of the silkworm is utilized to control the growth of the silkworm, increases the flexibility of silkworm breeding, cocoon purchasing and filature process, and has important implementation value.

Description

technical field [0001] The invention relates to a method for preparing a transgenic silkworm in the field of genetic engineering, in particular to a method for improving production traits by transgenic silkworm, and also relates to a method for controlling the development 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 one of the fundamental problems of silk industry science. 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 acq...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/34C12N15/866A01K67/04
Inventor 贡成良曹广力薛仁宇谢敏沈卫德
Owner SUZHOU UNIV
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