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Salmon's calcitonin gene synthesized by plant preference codon and its application

A salmon calcitonin, codon-preferred technology, applied to the expression of salmon calcitonin in plants, the field of the nucleic acid sequence of salmon calcitonin

Inactive Publication Date: 2003-01-15
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Before the publication of the present invention, there has not been any publication or report of the salmon calcitonin coding sequence mentioned in this patent application expressed in plants

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Synthesis of salmon calcitonin gene

[0049] 1. Gene synthesis

[0050] Since the activity of sCT is largely dependent on its conformation, amino acid residue substitution or deletion will also greatly change its activity (Yang Bin and Ding Zhenji, Chinese Journal of Pharmaceutical Sciences, 1995, 30: 643-646). According to the relationship between the activity and conformation of sCT, we made appropriate substitutions and deletions of its amino acid residues. For example, the valine (V) at position 8 of natural sCT is changed to glycine (G); the leucine (L) at position 16 is replaced with alanine (A), and the tyrosine (Y) at position 22 is deleted. ). Experiments have shown that the activity of the improved sCT is increased from 4500U / mg (natural sCT) before improvement to 8600U / mg after improvement. The adjusted primary structure of sCT is: CSNLSTCGLGKLSQEAHKLQTPRTNTGSGTP.

[0051] Based on the amino acid sequence of the primary structure of sCT and the principle ...

Embodiment 2

[0068] Sequence information and homology analysis of salmon calcitonin gene

[0069] The full length of the salmon calcitonin msCT1 gene of the present invention is 137 bp (including the linker), and the detailed sequence is shown in SEQ ID NO.4, wherein the open reading frame is located at nucleotides 22-117. According to the deduced amino acid sequence of the full-length salmon calcitonin, it has a total of 32 amino acid residues, a molecular weight of 3318.74, and a pI of 8.89. See SEQ ID NO.6 for the detailed sequence.

[0070] The full length of the salmon calcitonin msCT2 gene of the present invention is 140 bp (including the linker), and the detailed sequence is shown in SEQ ID NO.5, wherein the open reading frame is located at nucleotides 22-120. According to the deduced amino acid sequence of the full-length salmon calcitonin, it has a total of 33 amino acid residues, a molecular weight of 3375.79, and a pI of 8.89. See SEQ ID NO.7 for the detailed sequence.

[0071]...

Embodiment 3

[0073] Eukaryotic expression of salmon calcitonin in tobacco and tomato cells

[0074] Construction of Agrobacterium Strains Containing Expression Vectors of Target Genes (Salmon Calcitonin msCT1 and msCT2 Genes)

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Abstract

A salmon's calcitonin gene synthesized by plant preference codon, its fusion gene configurator, the recombinant expression carrier carrying said configurator, the plant cell converted by said expression carrier, the transgenic plant and its descendant generated by said converted cells for expressing said salmon's calcitonin, and the process for detecting the nucleic acid sequence and polypeptide of salmon's calcitonin in specimen are disclosed.

Description

technical field [0001] The invention relates to the fields of molecular biology, enzymology, physiology, genetic engineering and the like. Specifically, the present invention relates to a nucleic acid sequence of artificially synthesized salmon calcitonin (Modified Salmon Calcitonin, msCT) designed according to plant preferred codons for the first time. The present invention also relates to the expression and application of salmon calcitonin in plants. Background technique [0002] Calcitonin (calcitonin, CT) was discovered by Copp in 1962. It is a hormone with rapid and short-term calcium-lowering effect secreted by parafollicular cells (C cells) in the thyroid. The active CT is 32 peptides with a molecular weight of For 3500 Daltons, the 1st and 7th cysteines at the N-terminal form a disulfide ring, and the C-terminal is a proline (Pro) structure (Yang Shaohua and Chen Junjie, Progress in Biochemistry and Biophysics, 1996, 23:34-38). Hirsh first discovered and proved th...

Claims

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Application Information

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IPC IPC(8): C07H21/04C07K14/575C12N15/11C12N15/12C12N15/63C12N15/82C12P21/02C12Q1/68
Inventor 唐克轩赵凌侠孙小芬
Owner FUDAN UNIV
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