JPH07289261A - Gene of adhesive protein of mytilus edulis - Google Patents

Gene of adhesive protein of mytilus edulis

Info

Publication number
JPH07289261A
JPH07289261A JP6090266A JP9026694A JPH07289261A JP H07289261 A JPH07289261 A JP H07289261A JP 6090266 A JP6090266 A JP 6090266A JP 9026694 A JP9026694 A JP 9026694A JP H07289261 A JPH07289261 A JP H07289261A
Authority
JP
Japan
Prior art keywords
protein
gene
sequence
rna
mytilus edulis
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
JP6090266A
Other languages
Japanese (ja)
Other versions
JP3537487B2 (en
Inventor
Hiroshige Inoue
広滋 井上
Satoru Oudou
哲 王堂
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
KAIYO BIO TECH LAB
KAIYO BIO TECHNOL KENKYUSHO KK
Original Assignee
KAIYO BIO TECH LAB
KAIYO BIO TECHNOL KENKYUSHO KK
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by KAIYO BIO TECH LAB, KAIYO BIO TECHNOL KENKYUSHO KK filed Critical KAIYO BIO TECH LAB
Priority to JP09026694A priority Critical patent/JP3537487B2/en
Publication of JPH07289261A publication Critical patent/JPH07289261A/en
Application granted granted Critical
Publication of JP3537487B2 publication Critical patent/JP3537487B2/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Abstract

PURPOSE:To obtain the object gene useful for producing a peptide as a raw material for an adhesive effective even in the underwater or wet environment. CONSTITUTION:The objective gene of adhesive protein of Mytilus edulis codes an amino acid sequence of the formula. This gene is capable of producing the 2nd protein of an adhesive protein of Mytilus edulis by genetic engineering technique. The gene is obtained by the following processes: a poly A-RNA is prepared from the whole RNA obtained from the pedes of Mytilus edulis, and a double-stranded DNA is prepared using the poly A-RNA as template and then inserted into an appropriate vector; the resultant vector is transduced into an appropriate host followed by amplification to select a clone having the objective DNA and isolate the whole length of the 2nd protein cDNA, whose base sequence is then determined.

Description

【発明の詳細な説明】Detailed Description of the Invention

【0001】[0001]

【産業上の利用分野】本発明は水中や湿潤な環境で使用
できる接着剤の原料となるペプチドを組換えDNA技術
を用いて製造するために用いる遺伝子DNAに関する。
接着蛋白質をコードするDNAを組み込んだ組換え体D
NAを含む微生物や培養細胞を培養液中で培養し、該培
養物中に蓄積される該ポリペプチドを採集することによ
り、得られる該ペプチドは、接着剤の原料として広い用
途で利用されることが期待される。
BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a gene DNA used for producing a peptide as a raw material of an adhesive which can be used in water or a humid environment by using recombinant DNA technology.
Recombinant D incorporating a DNA encoding an adhesion protein
The peptide obtained by culturing a microorganism containing NA and cultured cells in a culture medium and collecting the polypeptide accumulated in the culture is widely used as a raw material of an adhesive. There is expected.

【0002】[0002]

【従来の技術】乾燥条件下で強い接着力を示す接着剤は
様々な種類のものが開発されている。そのうちの多くの
ものは一旦乾燥条件下で接着してしまえば湿潤環境にお
かれてもその強度を維持できる。しかし、湿潤な条件下
や水中で接着を開始した場合、有効な強度に達すること
ができる接着剤は存在しなかった。
2. Description of the Related Art Various types of adhesives have been developed which exhibit strong adhesiveness under dry conditions. Many of them can maintain their strength even in a humid environment once they are bonded under dry conditions. However, no adhesive was able to reach effective strength when the adhesion was initiated under moist conditions or in water.

【0003】ムラサキイガイは自己を良好な環境に固定
するために接着蛋白質を合成して自己を基物に接着させ
ることができる。この接着蛋白質には大まかには2種類
あり(Rzepecki, L.M., Hansen, K.M. and Waite, J.H.
Biological Bulletin 183:123-137, 1992)、両者の協
調作用により強固な接着が実現されることが想像され
る。一方の蛋白質(以降、第1蛋白質と称する)は主と
して数十個のAla-Lys-Pro-Ser-Tyr-Pro-Pro-Thr-Tyr-Ly
s という10アミノ酸の繰り返しにより構成される蛋白質
で、配列中のProおよびTyr残基はしばしば修飾されてそ
れぞれHydroxyprolineおよびDopa残基に変換されている
ことが知られており、Dopaのキノン架橋が接着に関与す
ることが推測されている(J.H.Wait, Int.J.Adhesion a
nd Adhesives, 7:9-14, 1987)。この蛋白質のcDNA
についてはすでに明らかにされ、その一部の配列を用い
て、微生物に作らせる方法がすでに報告されている(特
開平1-104180号公報)。もう一方の蛋白質(以降、第2
蛋白質と称する)はCys残基をその組成中に含む蛋白質
で、Cys残基のジスルフィド結合により、硬化する蛋白
質であると想像されるが、ムラサキイガイ足中に多量に
含まれることが知られているにもかかわらず、そのポリ
ペプチドの配列は一部の断片ペプチドのアミノ酸配列以
外は明らかにされていなかった。
The mussel can synthesize an adhesive protein to adhere itself to a substrate in order to fix itself in a favorable environment. There are roughly two types of this adhesion protein (Rzepecki, LM, Hansen, KM and Waite, JH
Biological Bulletin 183: 123-137, 1992), and it is envisioned that a strong bond is realized by the cooperative action of the two. One protein (hereinafter referred to as the first protein) is mainly composed of several tens of Ala-Lys-Pro-Ser-Tyr-Pro-Pro-Thr-Tyr-Ly.
It is known that the Pro and Tyr residues in the sequence are often modified and converted to Hydroxyproline and Dopa residues, respectively. (JHWait, Int.J.Adhesion a
nd Adhesives, 7: 9-14, 1987). CDNA of this protein
Has already been clarified, and a method for causing a microorganism to make a part of the sequence has already been reported (JP-A-1-104180). The other protein (hereinafter second
(It is called a protein) is a protein that contains Cys residues in its composition, and it is thought that it is a protein that hardens due to the disulfide bond of Cys residues, but it is known to be contained in large amounts in the mussel legs. Nevertheless, the sequence of the polypeptide was not revealed except for the amino acid sequences of some fragment peptides.

【0004】[0004]

【発明が解決しようとする課題】本発明は、遺伝子工学
の手法を用いてムラサキイガイ接着蛋白質の第2蛋白質
を生産すべく、その生産に係る遺伝子を提供することを
目的とする。
DISCLOSURE OF THE INVENTION An object of the present invention is to provide a gene involved in the production of a second protein of the mussel adhesion protein by using a genetic engineering technique.

【0005】[0005]

【課題を解決するための手段】ムラサキイガイ接着蛋白
質の第2蛋白質の配列を得るために、日本産のムラサキ
イガイより前記第2蛋白質をコードするcDNAを単離
した。日本産のムラサキイガイ(Mytilus galloprovinc
ialis)はチレニアイガイとも呼ばれる地中海系の種で
ある(Wilkins et al.Biol.J.Linnean Soc.,20:365-37
4,1983)。研究の結果、日本産ムラサキイガイの足から
抽出したmRNAから作成したcDNAライブラリーか
ら、6個のCys残基を一定の規則に基づいて含む32〜
34のアミノ酸残基からなるモチーフを、4〜9アミノ
酸残基のスペーサー領域を挟んで繰り返す構造から主と
して構成され、すでに知られているムラサキイガイ接着
蛋白質の第2蛋白質の断片配列と相同性のある配列を持
つcDNAを単離することに成功し、さらにその塩基配
列を決定して本発明を完成した。
[Means for Solving the Problems] In order to obtain the sequence of the second protein of the mussel adhesion protein, a cDNA encoding the second protein was isolated from Japanese mussels. Japanese Mussel (Mytilus galloprovinc)
ialis) is a Mediterranean species also called the Tyrene mussel (Wilkins et al. Biol. J. Linnean Soc., 20: 365-37).
4,1983). As a result of the research, a cDNA library prepared from mRNA extracted from Japanese Mussel mussel legs contained 6 Cys residues based on a certain rule.
A sequence mainly composed of a structure in which a motif consisting of 34 amino acid residues is repeated with a spacer region of 4 to 9 amino acid residues sandwiched therebetween, and a sequence homologous to the already known fragment sequence of the second protein of mussel adhesion protein. The present invention was completed by successfully isolating a cDNA having

【0006】即ち、本発明の第一は、配列番号2で表さ
れるアミノ酸配列をコードするムラサキイガイ接着蛋白
質遺伝子である。この遺伝子のDNA配列としては、配
列番号1で表される塩基配列を例示することができる。
That is, the first aspect of the present invention is a mussel adhesion protein gene encoding the amino acid sequence represented by SEQ ID NO: 2. As the DNA sequence of this gene, the base sequence represented by SEQ ID NO: 1 can be exemplified.

【0007】また、本発明の第二は、ミチルス・エドゥ
リス(Mytilus edulis)又はミチルス・ガロプロヴィンシ
アリス(Mytilus galloprovincialis) を起源とし、塩基
配列の長さが1.5Kbであり、配列番号3、4、又は5で
表されるアミノ酸配列を含むポリペプチドをコードする
ムラサキイガイ接着蛋白質遺伝子である。
The second aspect of the present invention originates from Mytilus edulis or Mytilus galloprovincialis and has a nucleotide sequence length of 1.5 Kb and SEQ ID NOs: 3 and 4. Or a mussel adhesion protein gene encoding a polypeptide containing the amino acid sequence represented by 5 or 5.

【0008】以下、本発明を詳細に説明する。本発明の
遺伝子を取得するための材料としては、日本産のムラサ
キイガイ(Mytilus galloprovincialis)を用いるのが
好ましいが、その他のムラサキイガイ、例えば、ミチル
ス・エドゥリス(Mytilus edulis)等を用いることもでき
る。本発明遺伝子は以下の手順で得ることができる。ま
ずムラサキイガイの足をチオシアン酸グアニジン等によ
り可溶化し、フェノール/クロロホルムによる抽出を行
い、イソプロパノールにより沈殿させることにより全R
NAを得ることができる。全RNAを得る方法はこの方
法に限定されるものではなく、LiCl沈殿法や塩化セ
シウム溶液に重層して遠心することによっても得られ
る。全RNAから、オリゴdTセルロースカラムを用い
てポリアデニル酸鎖を有するRNA(ポリA−RNA)
を調製する。このポリA−RNAを鋳型として逆転写酵
素を用いて2本鎖DNAを調製する。この2本鎖DNA
の合成はS1ヌクレアーゼ法やオカヤマ−バーグ法によ
り行ない得るが、市販のcDNA合成キットを用いて合
成することも可能である。次いで、得られたcDNAを
適当なベクターに挿入し、このベクターを適当な宿主に
導入して増幅させるとともに目的のDNAを持つクロー
ンを選択する。ベクターはλファージ由来の各種ベクタ
ーたとえばλgt10やλZAPIIなど、あるいはp
BR322等のプラスミドベクターを用いることができ
る。目的クローンの選択にはすでに知られているムラサ
キイガイ接着蛋白質の第2蛋白質の部分配列の一部に相
当するオリゴヌクレオチドを合成してプローブとして用
い、これに強く結合するクローンを選択すればよい。配
列の決定はサンガー法やマキサム−ギルバート法等の一
般的な方法によって決定できる。以上の手順により翻訳
開始コドンから終止コドン、さらにポリアデニル酸鎖付
加シグナルを含む第2蛋白質cDNAの全長を単離する
ことができる。
The present invention will be described in detail below. As a material for obtaining the gene of the present invention, Japanese mussel (Mytilus galloprovincialis) is preferably used, but other mussels such as Mytilus edulis can also be used. The gene of the present invention can be obtained by the following procedure. First, the mussels' legs are solubilized with guanidine thiocyanate, extracted with phenol / chloroform, and precipitated with isopropanol for total R
NA can be obtained. The method for obtaining total RNA is not limited to this method, and it can also be obtained by a LiCl precipitation method or a method in which a cesium chloride solution is layered and centrifuged. RNA having polyadenylic acid chains from total RNA using an oligo dT cellulose column (polyA-RNA)
To prepare. Double-stranded DNA is prepared using this poly A-RNA as a template and reverse transcriptase. This double-stranded DNA
Can be synthesized by the S1 nuclease method or the Okayama-berg method, but can also be synthesized by using a commercially available cDNA synthesis kit. Then, the obtained cDNA is inserted into an appropriate vector, this vector is introduced into an appropriate host for amplification, and a clone having the desired DNA is selected. Vectors are various vectors derived from λ phage, such as λgt10 and λZAPII, or p
A plasmid vector such as BR322 can be used. To select a target clone, an oligonucleotide corresponding to a part of the second protein partial sequence of the mussel adhesion protein, which is already known, is synthesized and used as a probe, and a clone that strongly binds to this may be selected. The sequence can be determined by a general method such as Sanger method or Maxam-Gilbert method. By the above procedure, the full length of the second protein cDNA including the translation initiation codon, the stop codon, and the polyadenylic acid chain addition signal can be isolated.

【0009】単離した配列は適当を発現ベクターに挿入
し、微生物や培養細胞に導入して発現させることによ
り、当該ペプチドを大量調製することが可能である。こ
の際、当該DNAはシグナル部分を含むため、当該ペプ
チドを宿主細胞外に分泌させることができる。また、シ
グナル部分を除去して適当なベクターに組み込んで用い
ることにより細胞内で生産させることも可能である。
The isolated sequence can be prepared in a large amount by inserting an appropriate sequence into an expression vector and introducing it into a microorganism or a cultured cell for expression. At this time, since the DNA contains a signal portion, the peptide can be secreted outside the host cell. It is also possible to produce intracellularly by removing the signal portion and incorporating it into an appropriate vector for use.

【0010】[0010]

【実施例】【Example】

〔実施例1〕 ムラサキイガイ足cDNAライブラリー
の作製 岩手県宮古市宮古湾で採集した殻長3−5cmのムラサ
キイガイ12個体の足をチオシアン酸グアニジン、クエ
ン酸ナトリウム、N−ラウリルザルコシン酸ナトリウ
ム、β−メルカプトエタノール等の溶液中で組織を機械
的に破砕し、フェノールおよびクロロホルムによる抽出
を行なって蛋白質等を除去したのち、イソプロパノール
を加えて沈殿させることにより全RNAを抽出し、オリ
ゴdTセルロースカラムに導通してポリアデニル酸鎖を
有するRNA(ポリA−RNA)を調製した。この操作
により約2μgのポリA−RNAが得られた。次にこの
ポリA−RNAを鋳型として逆転写酵素を用いて2本鎖
cDNAを調製した。この操作はアマシャム社のcDN
A合成キットを用いて添付のプロトコールにしたがって
行なった。ついで得られた2本鎖DNAにEcoRIア
ダプターを付加し、ファージベクターλgt10(アマ
シャム〔Amersham〕社製)に挿入した。この操作は、ア
マシャム社のcDNA合成システムを用いて添付のプロ
トコールにしたがって行なった。挿入の完了したファー
ジベクターは同キットに添付のインビトロパッケージン
グ溶液を用いて組換えDNAをファージ内に封入させ
た。封入の完了した組換えファージは、大腸菌NM51
4株(アマシャム〔Amersham〕社製)に感染させ、増幅
した。
[Example 1] Preparation of mussel pod leg cDNA library The legs of 12 mussels of 3-5 cm in shell length collected in Miyako Bay, Miyako City, Iwate Prefecture were treated with guanidine thiocyanate, sodium citrate, sodium N-lauryl sarcosinate, β -Mechanically disrupt the tissue in a solution such as mercaptoethanol, extract with phenol and chloroform to remove proteins, etc., and then add isopropanol to precipitate to extract total RNA, which is then applied to an oligo dT cellulose column. An RNA having a polyadenylic acid chain in a conductive state (poly A-RNA) was prepared. By this operation, about 2 μg of poly A-RNA was obtained. Next, double-stranded cDNA was prepared using this poly A-RNA as a template and reverse transcriptase. This operation is performed by Amersham's cDN
This was carried out using the A synthesis kit according to the attached protocol. Then, an EcoRI adapter was added to the obtained double-stranded DNA and inserted into the phage vector λgt10 (manufactured by Amersham). This operation was performed using the cDNA synthesis system of Amersham, according to the attached protocol. For the phage vector after the insertion, the recombinant DNA was encapsulated in the phage using the in vitro packaging solution attached to the kit. The encapsulated recombinant phage is Escherichia coli NM51.
Four strains (manufactured by Amersham) were infected and amplified.

【0011】〔実施例2〕 接着蛋白質cDNAを含む
組換えファージの選択 実施例1で得られた組換えファージを増幅させ、得られ
た5万個のプラークをナイロンメンブレン ハイボンド
N上に固定した。ついでムラサキイガイ第2接着蛋白質
の断片ペプチド配列の一部の相補鎖に相当するオリゴヌ
クレオチドプロープTA(T,C)AA(C,T)GA
CGACGACおよびTA(T,C)AA(C,T)G
ATGATGATをミリジェンサイクロンDNA合成機
により合成し、それぞれα32P−ATPによる端末ラベ
ルにより標識して、プラークハイブリダイゼーションを
行なった。その結果、プローブTA(T,C)AA
(C,T)GACGACGACでスクリーニングした2
0000クローンより、100個以上のプローブと結合
するプラークが得られた。これらのうち10個のプラー
クを任意に選び、挿入されているcDNAの長さをアガ
ロース電気泳動により調べて、最も長い挿入断片をもつ
ものについてファージベクターから制限酵素EcoRI
を用いて挿入断片を切り出し、プラスミドベクターpBlu
escriptIISK(+)(ストラタジーン〔Stratagene〕社製)
に挿入した。このプラスミドベクターをE.coli-Mgfp2-2
-7に導入した。E.coli-Mgfp2-2-7は、工業技術院生命工
学工業技術研究所に寄託番号FERM P-14288として寄託さ
れている(寄託日:平成6年4月21日)。
Example 2 Selection of Recombinant Phage Containing Adhesion Protein cDNA The recombinant phage obtained in Example 1 was amplified, and 50,000 plaques obtained were immobilized on a nylon membrane High Bond N. Next, an oligonucleotide probe TA (T, C) AA (C, T) GA corresponding to a part of the complementary strand of the fragment peptide sequence of the mussel second adhesion protein
CGACGAC and TA (T, C) AA (C, T) G
ATGATGAT was synthesized by a Milligen Cyclone DNA synthesizer, labeled with a terminal label of α 32 P-ATP, and subjected to plaque hybridization. As a result, the probe TA (T, C) AA
Screened with (C, T) GACGACGAC 2
Plaques that bind to 100 or more probes were obtained from 0000 clones. Of these, 10 plaques were arbitrarily selected, the length of the inserted cDNA was examined by agarose gel electrophoresis, and the one having the longest insert fragment was digested with the restriction enzyme EcoRI from the phage vector.
The insert fragment was cut out using the plasmid vector pBlu
escriptIISK (+) (manufactured by Stratagene)
Inserted in. Use this plasmid vector for E. coli-Mgfp2-2
-7 introduced. E.coli-Mgfp2-2-7 has been deposited at the Institute of Biotechnology, Institute of Industrial Science and Technology under the deposit number FERM P-14288 (deposit date: April 21, 1994).

【0012】〔実施例3〕 接着蛋白質遺伝子の配列決
定 実施例2で得られた挿入断片の配列をアプライドバイオ
システムズ社製373ADNAシーケンサーおよびシー
ケンシングキットを用いてDNA配列を決定した。その
結果、この挿入断片が接着蛋白質の全長を含むDNA配
列であることが判明した。得られた接着蛋白質遺伝子
は、配列番号1に示す通り、翻訳開始コドンから翻訳終
了コドンまで473アミノ酸配列をコードする全長15
15bpの配列であり、473アミノ酸のうち最上流か
ら17残基がシグナルペプチド、続く31残基が非繰り
返し領域であり、その下流に6個のCys残基を一定の規
則に基づいて含む32−34個のアミノ酸残基からなる
モチーフ、すなわち配列番号3、4又は5で表されるア
ミノ酸配列を4−9アミノ酸残基のスペーサー領域を挟
んで繰り返す繰り返し領域、さらに下流には13アミノ
酸残基の非繰り返し領域が存在した。翻訳終了コドンの
下流側の非翻訳領域にはポリアデニル酸鎖付加シグナル
ATTAAAが存在し、そのさらに下流にポリアデニル
酸鎖が存在した。
[Example 3] Sequencing of Adhesion Protein Gene The DNA sequence of the insert fragment obtained in Example 2 was determined using an Applied Biosystems 373A DNA sequencer and sequencing kit. As a result, it was revealed that this insert fragment was a DNA sequence containing the entire length of the adhesion protein. As shown in SEQ ID NO: 1, the resulting adhesion protein gene has a total length of 15 amino acids encoding a 473 amino acid sequence from the translation initiation codon to the translation termination codon.
It is a sequence of 15 bp, 17 residues from the most upstream of 473 amino acids are a signal peptide, the following 31 residues are a non-repetitive region, and 6 Cys residues are contained downstream thereof based on a certain rule. A motif consisting of 34 amino acid residues, that is, a repeating region in which the amino acid sequence represented by SEQ ID NO: 3, 4 or 5 is repeated with a spacer region of 4-9 amino acid residues in between, and further downstream is 13 amino acid residues. There were non-repeating regions. A polyadenylic acid chain addition signal ATTAAA was present in the untranslated region downstream of the translation termination codon, and a polyadenylic acid chain was present further downstream thereof.

【0013】[0013]

【発明の効果】本発明はムラサキイガイ接着蛋白質遺伝
子を提供する。本発明の遺伝子から作られる蛋白質は、
接着剤の原料として極めて有用である。
INDUSTRIAL APPLICABILITY The present invention provides a mussel adhesion protein gene. The protein made from the gene of the present invention is
It is extremely useful as a raw material for adhesives.

【0014】[0014]

【配列表】[Sequence list]

【0015】配列番号 1 配列の長さ:1515 配列の型 :核酸 鎖の数 :2本鎖 トポロジー:直鎖状 配列の種類:mRNA to cDNA 起源 生物名 :ムラサキイガイ(Mytilus galloprovincial
is) 配列
SEQ ID NO: 1 Sequence length: 1515 Sequence type: Nucleic acid Number of strands: Double strand Topology: Linear Sequence type: mRNA to cDNA Origin organism name: Blue mussel (Mytilus galloprovincial)
is) array

【0016】配列番号 2 配列の長さ:473 配列の型 :アミノ酸 トポロジー:不明 配列の種類:タンパク質 起源 生物名 :ムラサキイガイ(Mytilus galloprovincial
is) 配列
SEQ ID NO: 2 Sequence length: 473 Sequence type: Amino acid Topology: Unknown Sequence type: Protein Origin organism name: Blue mussel (Mytilus galloprovincial)
is) array

【0017】配列番号 3 配列の長さ:32 配列の型 :アミノ酸 トポロジー:不明 配列の種類:ペプチド 起源 生物名 :ムラサキイガイ(Mytilus galloprovincial
is) 配列
SEQ ID NO: 3 Sequence length: 32 Sequence type: Amino acid Topology: Unknown Sequence type: Peptide Origin Biological name: Mytilus galloprovincial
is) array

【0018】配列番号 4 配列の長さ:33 配列の型 :アミノ酸 トポロジー:不明 配列の種類:ペプチド 起源 生物名 :ムラサキイガイ(Mytilus galloprovincial
is) 配列
SEQ ID NO: 4 Sequence length: 33 Sequence type: Amino acid Topology: Unknown Sequence type: Peptide Origin Biological name: Mytilus galloprovincial
is) array

【0019】配列番号 5 配列の長さ:34 配列の型 :アミノ酸 トポロジー:不明 配列の種類:ペプチド 起源 生物名 :ムラサキイガイ(Mytilus galloprovincial
is) 配列
SEQ ID NO: 5 Sequence length: 34 Sequence type: Amino acid Topology: Unknown Sequence type: Peptide Origin organism name: Blue mussel (Mytilus galloprovincial)
is) array

───────────────────────────────────────────────────── フロントページの続き (51)Int.Cl.6 識別記号 庁内整理番号 FI 技術表示箇所 C12R 1:19) ─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. 6 Identification code Office reference number FI technical display location C12R 1:19)

Claims (3)

【特許請求の範囲】[Claims] 【請求項1】 配列番号2で表されるアミノ酸配列をコ
ードするムラサキイガイ接着蛋白質遺伝子。
1. A mussel adhesion protein gene encoding the amino acid sequence represented by SEQ ID NO: 2.
【請求項2】 DNA配列が配列番号1で表される請求
項1記載のムラサキイガイ接着蛋白質遺伝子。
2. The mussel adhesion protein gene according to claim 1, wherein the DNA sequence is represented by SEQ ID NO: 1.
【請求項3】 ミチルス・エドゥリス(Mytilus edulis)
又はミチルス・ガロプロヴィンシアリス(Mytilus gallo
provincialis) を起源とし、塩基配列の長さが1.5Kbで
あり、配列番号3、4、又は5で表されるアミノ酸配列
を含むポリペプチドをコードするムラサキイガイ接着蛋
白質遺伝子。
3. Mytilus edulis
Or Mytilus gallo
mussel adhesion protein gene originating in Provincia lis) and having a nucleotide sequence of 1.5 Kb and encoding a polypeptide containing the amino acid sequence represented by SEQ ID NO: 3, 4 or 5.
JP09026694A 1994-04-27 1994-04-27 Purple mussel adhesion protein gene Expired - Fee Related JP3537487B2 (en)

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Application Number Priority Date Filing Date Title
JP09026694A JP3537487B2 (en) 1994-04-27 1994-04-27 Purple mussel adhesion protein gene

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US7910699B2 (en) 2005-06-10 2011-03-22 Basf Se Cysteine-depleted hydrophobin fusion proteins, their production and use thereof
US8038740B2 (en) 2005-10-12 2011-10-18 Basf Se Use of proteins as an antifoaming constituent in fuels
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KR100868047B1 (en) * 2002-08-13 2008-11-10 주식회사 포스코 ADHESIVE PROTEIN Mgfp-5 ISOLATED FROM MUSSEL AND METHOD OF PRODUCING SAME
US7892788B2 (en) 2005-02-07 2011-02-22 Basf Se Hydrophobin fusion products, production and use thereof
WO2006103225A1 (en) * 2005-03-31 2006-10-05 Basf Aktiengesellschaft Use of polypeptides in the form of adhesive agents
JP4772110B2 (en) * 2005-03-31 2011-09-14 ビーエーエスエフ ソシエタス・ヨーロピア Use of polypeptides as adhesion promoters
US8859106B2 (en) 2005-03-31 2014-10-14 Basf Se Use of polypeptides in the form of adhesive agents
US7799741B2 (en) 2005-04-01 2010-09-21 Basf Se Drilling mud containing hydrophobin
US8535535B2 (en) 2005-04-01 2013-09-17 Basf Se Use of hydrophobin as a phase stabilizer
US7910699B2 (en) 2005-06-10 2011-03-22 Basf Se Cysteine-depleted hydrophobin fusion proteins, their production and use thereof
US8038740B2 (en) 2005-10-12 2011-10-18 Basf Se Use of proteins as an antifoaming constituent in fuels
US8096484B2 (en) 2006-08-15 2012-01-17 Basf Se Method for the production of dry free-flowing hydrophobin preparations

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