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Application of sheep KAP gene in mechanical property improvement and dyeing property improvement of natural silk, and recombinant expression vector of the KAP gene

A technology for expressing vectors and mechanical properties, applied in the direction of plant gene improvement, using vectors to introduce foreign genetic material, applications, etc., can solve problems such as the gap in comprehensive performance, achieve strong mechanical properties, good dyeing performance, and overcome poor dyeing performance.

Inactive Publication Date: 2014-07-09
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unfortunately, so far, although the mechanical properties of silk obtained in the study of improving the mechanical properties of silk by using spider silk have been improved to a certain extent, the comprehensive properties such as taking into account toughness, rigidity, elasticity, etc. are still somewhat different from those of natural spider silk. However, there is no report on the dyeing performance of transgenic cocoon silk

Method used

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  • Application of sheep KAP gene in mechanical property improvement and dyeing property improvement of natural silk, and recombinant expression vector of the KAP gene
  • Application of sheep KAP gene in mechanical property improvement and dyeing property improvement of natural silk, and recombinant expression vector of the KAP gene
  • Application of sheep KAP gene in mechanical property improvement and dyeing property improvement of natural silk, and recombinant expression vector of the KAP gene

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Sequence optimization of wool keratin-associated proteins (KAP)

[0025] According to the preference of endogenous gene expression sequence codons in the silkworm genome sequence data, the present invention is derived from sheep KAP Gene sequences (gene accession numbers: GI: 238637219 and GI: 57164397) were optimized and designed, and the optimized nucleotide sequences such as SEQ ID NO.1 and SEQ ID NO.2 were named KAP4.3 and KAP5.4, respectively. Its amino acid sequence is shown in SEQ ID NO.3 and SEQ ID NO.4.

[0026] 2. Construction of Bombyx mori transgene recombination vector

[0027] According to the requirements of the currently known and public transgenic technology based on piggyBac transposable vectors in silkworms, the silkworm transgenic recombinant vector pBac{3×P3-DsRed; FibH-KAP4.3-LBS} was constructed ( figure 1 A) and pBac{3×P3-DsRed; FibH-KAP5.4-LBS} ( figure 1 B). Its detailed preparation method is as follows: the artificially synthesized KAP...

Embodiment 2

[0029]The commercial diapause silkworm strain 871 was used as the raw material, and the parental silkworm eggs were treated with low-temperature acceleration at 16°C to relieve the diapause of the offspring silkworm eggs; then 10-15 nL of the recombinant vector pBac{3×P3 -DsRed; FibH-KAP4.3-LBS} and the helper plasmid pHA3PIG were injected into 247 G0 silkworm eggs released from diapause, sealed with non-toxic glue and placed in an environment at 25°C and 85% relative humidity Incubating in the middle of greening, 177 G0 generation silkworms were hatched, and then the silkworms were raised with mulberry leaves until they turned into moths, and 36 G0 generation silkworms were obtained, and a total of 22 circles of G1 generation silkworms were obtained by backcrossing or selfing the obtained silkworm moths The eggs were observed with an Olympus? motorized macroscopic fluorescence microscope, and one moth circle emitting red fluorescence was obtained after screening, and a total o...

Embodiment 3

[0031] The commercial diapause silkworm strain 871 was used as the raw material, and the parental silkworm eggs were treated with low-temperature acceleration at 16°C to relieve the diapause of the offspring silkworm eggs; The mixture of DsRed; ibH-KAP5.4-LBS} and the helper plasmid pHA3PIG was injected into 234 G0 silkworm eggs released from diapause, sealed with non-toxic glue and placed in an environment at 25°C and 85% relative humidity Accelerate the hatching and hatch to obtain 126 G0 generation silkworms, and then raise the silkworms with mulberry leaves until they become moths, and obtain 17 G0 generation silkworm moths, and obtain 10 rounds of G1 generation silkworm eggs by backcrossing or selfing the obtained silkworm moths , observed with an Olympus? motorized macroscopic fluorescent microscope, and obtained 1 red fluorescent moth circle after screening, and obtained 6 positive transgenic silkworms with red fluorescent eyes, and the transformation efficiency was 4.5%...

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Abstract

The invention discloses an application of sheep KAP gene in mechanical property improvement and dyeing property improvement of natural silk, and a recombinant expression vector of the KAP gene. A wool keratin associated protein (KAP) is subjected to recombinant expression in silkworms, and then the KAP and fibroin that is the main part of bave are subjected to fusion expression. The KAP protein can be uniformly and stably distributed in the fibroin in fibroin secretion processes of silk glands, thus improving the mechanical property and the dyeing property of the natural silk, and laying good theoretical and practical bases for large-scale production of novel medical and military materials and other spun silk materials with high added value by utilization of the bave.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an application of sheep KAP gene in improving silk mechanical properties and dyeing properties and a recombinant expression vector thereof. Background technique [0002] China is a big silk country and the birthplace of the ancient Silk Road. The silkworm is one of the most successful and longest economic insects utilized by humans in the world at present. The domestication and breeding of the silkworm have a history of about 5,000 years. Silkworm cocoons obtained by sericulture are still one of the important textile industry raw materials so far. As a textile material, although silk has the advantages of good hygroscopicity and strong warmth retention, it also has friction strength and buckling strength inferior to synthetic fibers, spun silk materials are prone to wrinkling and not strong enough, and silk fabrics are prone to yellowing and fading after washing and su...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/47C12N15/85
Inventor 赵爱春张阳龙定沛鲁成向仲怀
Owner SOUTHWEST UNIVERSITY
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