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Method for production of tracp5b

A coding and silkworm technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of failure to establish production methods, unable to reproduce TRACP5b post-translational modification, etc.

Active Publication Date: 2011-08-10
NITTO BOSEIKI CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The reason for this is that the specific post-translational modification in the production of TRACP5b cannot be reproduced in these systems (Non-Patent Document 4)
In addition, the present inventors tried to prepare TRACP5 and perform enzymatic treatment with cathepsin K etc. in vitro, but failed to establish a production method of an enzyme protein that is stable from an industrial point of view

Method used

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  • Method for production of tracp5b
  • Method for production of tracp5b
  • Method for production of tracp5b

Examples

Experimental program
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Embodiment

[0236] Hereafter, although an Example demonstrates this invention more concretely, this invention is not limited to these Examples.

[0237] Materials and Methods

[0238] [carrier build]

[0239] In the present invention, the plasmid vector pBMCSUASsigTRACP ( figure 1 and SEQ ID NO: 1). In this vector for preparing recombinant silkworm, downstream of the promoter UAS (which promotes gene expression in the presence of the yeast transcriptional regulator GAL4), there is a human TRACP gene. In addition, as a marker gene for identifying recombinant silkworm cocoons, the vector has a green fluorescent protein gene 3xP3GFP linked to a promoter capable of promoting expression in ocelli of embryos, compound eyes of moths, and nerve-derived tissues (Horn, C ., and E.A. Wimmer, (2000) Dev Genes Evol 210:630-637; Murizio et al. (1994) Protein Science, 3:1476-1484).

[0240] [Preparation of recombinant silkworm and establishment of recombinant protein expression strain]

[0241] ...

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Abstract

A silkworm is produced, which has: (i) operably linked DNA encoding a transcription regulation factor, which is located downstream to a promoter for DNA encoding a protein which is expressed in a silk-gland-specific manner; and (ii) operably linked DNA encoding a tartrate resistant acid phosphatase 5b (TRACP5b), which is located downstream to a target promoter for the transcription regulation factor. As a result, it is found that the silkworm produces a TRACP5 having an activity. This means that a TRACP5 produced from a silk gland of the silkworm undergoes a processing at a silk gland in the same manner as in the processing at a site wherebone resorption occurs.

Description

technical field [0001] The present invention relates to a method for producing tartrate-resistant acid phosphatase 5b (TRACP5b) using silkworm. Furthermore, the present invention relates to silkworms producing TRACP5b. Background technique [0002] Acid phosphatase in serum was separated into 6 bands ranging from 0 to 5 from the origin by polyacrylamide gel electrophoresis. The fifth of these bands shows tartrate resistance and is called Band 5 tartrate resistant acid phosphatase (TRACP 5: Tartrate Resistant Acid Phosphatase 5). [0003] The fifth band was further divided into two bands in acidic disc electrophoresis, called TRACP5a and TRACP5b respectively. TRACP5a is an enzyme derived from platelets or other components, and its blood levels are unchanged. On the other hand, since the blood level of TRACP5b varies with bone resorption, it is considered to be derived from osteoclasts. In the development of specific inhibitors, activators or modulators of TRACP5b, a large...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/16C12Q1/42C12N15/09G01N33/573
CPCC12N9/16C12N15/09C12Q1/42G01N33/573
Inventor 清川岩大桥建也三浦俊英片山胜博田村俊树小林功瀬筒秀树
Owner NITTO BOSEIKI CO LTD