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Identification of Functional Protein Regions Through Chimeric Protein Construction
Published on: January 8, 2019
13種類の異なる筋肉タンパク質のための新しいトロポニンTとcDNAクローン,ショットガン配列解析で発見された
Nature
|April 21, 1983
まとめ
研究者らは,補完的なDNA (cDNA) のライブラリをシーケンシングすることによって,新しい筋肉タンパク質の変種を特定しました. この方法は,豊富な筋肉タンパク質のクローンを効率的に発見し,新しい配列変異を発見し,筋肉タンパク質の多様性を理解するのに役立ちます.
科学分野:
- 分子生物学は分子生物学である.
- プロテオミクス プロテオミクスは,プロテオミクスの
- ゲノミクスゲノミクスとは
背景:
- 19種類の豊富な筋肉関連タンパク質のアミノ酸配列が確立されています.
- これらのタンパク質のmRNAレベルは,約0.4%またはそれ以上であると仮定した.
- 理論的分析は,タンパク質の識別のためのcDNAライブラリスクリーニングを提案しました.
研究 の 目的:
- 豊富な筋肉タンパク質のための特定のcDNAクローンを識別する.
- トロポニンを含む主要な筋肉タンパク質のアミノ酸配列変異を発見すること.
- 新しい筋肉タンパク質のイソタイプを特徴付けるために.
主な方法:
- ウサギの筋肉のcDNAライブラリでランダムに選択されたクローンからのcDNA挿入の配列決定.
- 178個のランダムに選択されたクローン内の約20,000個のヌクレオチドの決定.
- ニュクレオチド配列の比較分析により,変異と新しい同型を特定する.
主要な成果:
- 19の既知の筋肉特異タンパク質のうち13のクローンを特定した.
- 新しい同型を含む2種類のトロポニンTをコードする配列を発見した.
- トロポニンCの2つの既知の骨格筋同型体のシーケンスが確認されました.
- 新しいトロポニンT同型の核酸配列の有意な差異が観察されました.
結論:
- cDNAライブラリのランダムシーケンシングは,筋肉タンパク質のクローンを識別するための効果的な方法です.
- このアプローチは,新しいタンパク質同型型および配列変異を成功裏に発見することができます.
- この発見は,筋肉のタンパク質の多様性と遺伝子発現のより深い理解に貢献します.
関連する概念動画
From DNA to Protein
The flow of genetic information in cells from DNA to mRNA to protein is described by the central dogma, which states that genes specify the sequence of mRNAs, which in turn specify the sequence of amino acids making up all proteins. The decoding of one molecule to another is performed by specific proteins and RNAs. Because the information stored in DNA is so central to cellular function, it makes intuitive sense that the cell would make mRNA copies of this information for protein synthesis...
Transfer RNA Synthesis
One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
tRNA Activation
Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
Ribosome Profiling
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Peptide Identification Using Tandem Mass Spectrometry
Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...

