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Branchiostoma belcheri actin gene promoter and application thereof

A technology for actin and amphioxus, which is applied in the introduction of foreign genetic material, DNA/RNA fragments, recombinant DNA technology using vectors, etc., can solve problems such as research gaps, and achieve the effect of high start-up efficiency

Inactive Publication Date: 2012-07-11
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been well applied in model animals such as transgenic zebrafish and mice, but the study on the promoter of the amphioxus actin gene is still blank

Method used

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  • Branchiostoma belcheri actin gene promoter and application thereof
  • Branchiostoma belcheri actin gene promoter and application thereof
  • Branchiostoma belcheri actin gene promoter and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Acquisition of Actin Gene Promoter Sequence of Amphiprion

[0031] 1. The actin gene family identification and EST data processing of Amphiprion

[0032] Using the actin gene sequence of Florida Amphioxus (19, Wei, Y., H., Zhang, Y., J., Chen, Y., Mao, B., Y., 2009. Expanssion of the The Actin Gene Family in amphioxue. Zoological Research, 30(5): 473-479) used as bait to locate all Amphioxue actin genes from the genome just released from Sun Yat-sen University through homology sequence analysis BlastN program. A total of 33 actin genes were found. These genes were classified through evolutionary analysis, and the sequence alignment of actin protein was completed by CLUSTAL_W program (20, Thompson, J., D., Higgins, D., G., Gilson, T., J., 1994 .CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucl. Acids Res. (1994) 22(22): 4673-4680), the compariso...

Embodiment 2

[0060] Example 2 Amphioxus leucocephalus heat shock protein 70 gene promoter

[0061] 1. Construction of recombinant expression plasmid

[0062] The PCR products of the amplified two promoter regions were digested with HindIII and Not I, and loaded into the LacZ (the vector has no promoter, gifted by Professor You Zhikai, Academia Sinica, Taiwan) that has undergone the same double digestion treatment. The DH5α strain was transformed, and the positive clones were obtained after PCR and restriction enzyme digestion, and sequenced to prove that the inserted fragments were correct. Similarly, the enhancer fragments and random fragments obtained by PCR were treated with HindIII single restriction enzyme digestion, and then loaded into the single enzyme digestion treated by HindIII The obtained LacZ reporter vector inserted into 129bp was transformed, digested with restriction enzymes, and sequenced to obtain a positive clone, and finally the function was verified. A total of 4 LacZ rec...

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Abstract

The invention discloses a Branchiostoma belcheri actin gene promoter and application thereof, relate to a Branchiostomactin gene and provides the Branchiostoma belcheri actin gene promoter and application thereof to the regulation of expression of the Branchiostoma belcheri gene. The Branchiostoma belcheri actin gene promoter can effectively regulate the expression of a downstream gene; a cloned and separated minimal promoter can be applied to study on functions of Branchiostoma transcription regulating elements, such as an enhancer, while a basic promoter of the acting gene can be applied tostudy on interested gene functions at different times and in different spaces; and a promoter-mediated target gene is integrated into a Branchiostoma genome to obtain a transgenic Branchiostoma, so that target genes in different developmental stages or at different parts of the Branchiostoma can be studied.

Description

Technical field [0001] The present invention relates to amphioxus actin gene, and particularly relates to amphorae whites actin gene promoter and its application. Background technique [0002] Amphioxus belongs to the Cephalochordate of Chordate. It is at an important transitional stage in the evolution of invertebrates to vertebrates. It is the most similar to the direct ancestors of vertebrates in existing organisms. One of the taxa. Because of its unique evolutionary position, simple structure and translucent body, the genome is only 1 / 6 of that of humans, showing the characteristics of the genome of vertebrate ancestors before gene multiplication (1, Balczarek, KA, Lai, ZC, Kumar, S ., Evolution of functional diversification of the pair box (Pax) DNA-binding domains. Molecular. Biology and Evolution [J]. 1997. (14), 829-842.). With the deepening of research on Amphioxus, the whole genome sequencing of Florida Amphioxus (Branchiostoma flloridae) and White Amphioxus (Branchio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/85
Inventor 冯俊王义权李光刘欣
Owner XIAMEN UNIV
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