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Sebastiscus marmoratus CYP1B1 gene full-length sequence and cloning method thereof

A cloning method, the technology of the brown catfish, is applied to the full length of the marine biomarker CYP1B1 gene and its cloning field, which can solve the problems of few CYPs subgroup genes and no full-length reports of the brown pomfret

Inactive Publication Date: 2015-03-25
XIAMEN UNIV
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Benefits of technology

This patented method allows researchers to study how certain proteins are involved with harmful substances that can be found on various parts of living creatures such as humans or animals. By studying this process it helps identify which specific compounds may cause damage when exposed to these chemical agents.

Problems solved by technology

The technical problem addressed by this patents relates to understanding how algae metabolizing cycling reactions play crucial roles during natural environments like streams or lakes that contain harmful chemical compounds from their surroundings. This knowledge could help identify potential sources of contamination through analysis of these chemosignals.

Method used

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  • Sebastiscus marmoratus CYP1B1 gene full-length sequence and cloning method thereof
  • Sebastiscus marmoratus CYP1B1 gene full-length sequence and cloning method thereof
  • Sebastiscus marmoratus CYP1B1 gene full-length sequence and cloning method thereof

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Embodiment Construction

[0020] The following examples will further illustrate the present invention.

[0021] In the implementation process, the technical solution of the present invention can adopt the following concrete steps:

[0022] 1. Collection of animal tissue and organ samples

[0023] The fresh brown pomfret liver was collected, the blood on the surface was washed away with normal saline, and the total RNA was extracted immediately.

[0024] 2. Extraction of total RNA of scorpionfish

[0025] Total RNA was extracted according to the instructions of the Trizol kit, and the specific method was as follows:

[0026] a) Put the liver organ (about 100 mg) of Scorpene scorpionfish into a mortar, add 1 mL Trizol, vibrate with a shaker, let stand on ice for 5 min; centrifuge at 12000 r / min for 5 min at 4°C.

[0027] b) Take 700 μL of the supernatant, add 0.2 mL of chloroform, shake and mix (about 45 s), let stand at room temperature for 5 min; centrifuge at 12,000 r / min at 4°C for 15 min, after c...

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Abstract

The invention discloses a sebastiscus marmoratus CYP1B1 gene full-length sequence and a cloning method thereof, and relates to the field of marine genetic engineering. The cloning method comprises the following steps: extracting total RNA by using liver tissues of sebastiscus marmoratus, performing reverse transcription amplification to obtain 5'-end and 3'-end unknown sequences by adopting RACE, and splicing to obtain full-length cDNA sequences of the unknown sequences. By adopting the cloning method, a full-length CDS sequence of a CYP1B1 gene namely the only member of a CYP1B subfamily in the sebastiscus marmoratus is obtained, the blank of a P450 CYP1B gene family of the sebastiscus marmoratus is complemented, sea area pollution conditions can be indicated more accurately, and the problems of protein expression and the like of marine lives in polluted sea areas are explored more deeply. By virtue of treatment and preparation of materials, RNA extraction, synthesis of 5-end and 3-end cDNA, an RACE technology and the like, the CYP1B1 gene is obtained, wherein the cDNA full length of the gene is 3056bp, a gene open reading frame is 1724-2995bp, and totally 422 amino acids are coded.

Description

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Claims

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Application Information

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Owner XIAMEN UNIV
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