JP2009155295A - Larval settlement-inducing protein of balanus amphitrite - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a protein able to induce larval settlement of Balanus amphitrite in a wide range, an attractant for larvae of Balanus amphitrite using the protein, and a method for attracting the larvae of Balanus amphitrite. <P>SOLUTION: The protein is obtained from an extract of grown Balanus amphitrite and has an amino acid sequence comprising a specific sequence on the N terminal side and 32 kDa molecular weight. The protein has expressed a function inducing settlement of the larvae of Balanus amphitrite in a state in which the protein is dissolved in sea water and dispersed. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a protein that induces the establishment of larvae of the vertical barnacle. More specifically, the present invention relates to a pheromone-like protein that induces the establishment of the larvae of the attachment stage of the vertical barnacle, an attractant for the vertical barnacle larvae, an attracting substrate and an attracting method using the protein.

  Barnacles are typical fouling organisms that cause damage by adhering to offshore structures, thermal and nuclear power plant cooling water systems, ships, fishnets, and the like. For example, when a barnacle adheres to the cooling water channel system of a thermal power / nuclear power plant, the flow rate of the cooling water decreases due to an increase in flow resistance, and the cooling efficiency of the condenser decreases. In addition, when the barnacle adheres to the condenser pipe or the peeled barnacle is clogged, the pipe wall is corroded. Therefore, at each power plant, various control measures are taken to remove the attached barnacles mechanically or to suppress the attachment of barnacles. For example, the adhesion of barnacle larvae is suppressed by applying an antifouling paint to a cooling water channel system, a condenser pipe, or the like, or injecting chlorine (Non-patent Document 1).

  However, in the current situation, antifouling paints and methods using chlorine cannot sufficiently prevent the attachment of barnacle larvae. In addition, the use of antifouling paints and chlorine may adversely affect the environment.

  Therefore, a technique has been proposed to prevent the attachment of barnacles without adversely affecting the environment by using the induction-induced pheromone, which is one of the factors affecting the growth of barnacles. (Non-patent document 2). This anti-adhesion technology utilizes a sugar chain-binding protein complex (hereinafter referred to as SIPC) composed of 76, 88 and 99 kDa subunits that induces the establishment of vertebrate larvae larvae by adsorbing to a substrate or the like. This controls the adhesion of the vertical barnacle larvae.

By placing the base or the like having the SIPC adsorbed on the surface in the seawater, it is possible to induce the growth of the vertical barnacle larvae in the vicinity of the surface of the base and attach it to the surface of the base. Therefore, by changing the setting area of the vertical barnacle larvae, it is possible to suppress the attachment of the vertical barnacle larvae to marine structures, cooling water systems of thermal power / nuclear power plants, ships, fish nets, and the like.
Sakaguchi Isamu (2003). Prospects for antifouling organisms at power plants. Sessile Organisms 20 (1), 15-19. K.Matsumura, M.Nagano and N.Fusetani (1998) .Purification of a larval settlement-inducing protein complex (SIPC) of the barnacle, Balanus amphitrite.J. Exp.Zool., 281,12-20.

  Since the SIPC has a function of inducing the formation of the vertical barnacle larvae while adsorbed on the substrate or the like, the SIPC is required to be adsorbed on the substrate and used. Therefore, the range that can induce the establishment of the vertical barnacle larvae is limited to the vicinity of the substrate surface.

  However, if the range that can induce the larvae of the vertical barnacle larvae is limited to the vicinity of the surface of the basement, there is a problem that the larvae of the vertical barnacle larvae existing in a wide range of seawater cannot be sufficiently induced.

  Therefore, an object of the present invention is to provide a protein capable of inducing a wide range of epiphysis of a vertical barnacle larvae.

  Another object of the present invention is to provide an attractant, an attracting base material, and an attracting method capable of attracting a long-distance barnacle larva existing in a wide range in seawater to a desired position.

  In order to solve such a problem, the present inventor conducted intensive research.As a result, a protein having a specific amino acid sequence and a molecular weight among a plurality of proteins contained in the extract of the adult vertical barnacle is found in seawater. It has been found that it exhibits the function of inducing the growth of the vertical barnacle larvae in a dissolved and dispersed state. In addition, we investigated the behavior of the vertical barnacle larvae when this protein concentration gradient was formed in seawater, and found that it was possible to attract the vertical barnacle larvae to the higher concentration side of this concentration gradient. Based on these findings, the present invention has been achieved.

  Based on this finding, the protein according to claim 1 is obtained from an adult extract of Balanus amphitrite, has the amino acid sequence of SEQ ID NO: 1 on the N-terminal side, and has a molecular weight of 32 kDa It is.

  This protein is a novel protein having a function of inducing the growth of the vertical barnacle larvae when dissolved and dispersed in seawater.

  The attractant for the vertical barnacle larvae according to claim 2 is the one in which the protein according to claim 1 is supported on a substrate surface or a carrier. Moreover, the attractant for the vertical barnacle larvae according to claim 3 is the protein according to claim 1 formed into a solid form.

  According to the attractant for the vertical barnacle larvae according to claim 2 and 3, when the attractant is placed in seawater, the protein gradually elutes from the attractant and diffuses over a wide area, and the widespread of the vertical barnacle larvae is widespread. Induced by Further, when the protein is gradually eluted from the attractant, a concentration gradient between the attractant and the protein is formed, and the vertical barnacle larvae are attracted around the attractant.

  In the attracting base material for the vertical barnacle larvae according to claim 4, the protein according to claim 1 is attached to the surface of the base material.

  According to the attracting base material for the vertical barnacle larvae according to claim 4, when the attracting base material is placed in seawater, the protein gradually elutes from the surface of the attracting base material and diffuses over a wide area. Life is induced extensively. In addition, the protein gradually elutes from the attracting agent, thereby forming a protein concentration gradient centering on the attracting base material, and attracting the vertical barnacle larvae to the attracting base material.

  A method for attracting a vertical barnacle larvae according to claim 5 is to continuously supply the protein according to claim 1 to an attracting target area of the vertical barnacle larva to form a protein concentration gradient centering on the attracting target area. ing. Therefore, the protein diffuses over a wide area in the seawater, and the establishment of the vertical barnacle larvae is induced over a wide area, and the vertical barnacle larvae are attracted to the attracting target region having a high protein concentration.

  According to the protein of the first aspect, it exhibits a function of inducing the growth of the vertical barnacle larvae in a state of being dissolved and dispersed in seawater, so that it is possible to induce the widespread propagation of the vertical barnacle larvae.

  According to the attractant for the vertical barnacle larvae according to claims 2 and 3, it is possible to induce the widespread growth of the vertical barnacle larvae in a wide range and attract them to the periphery of the attractant. Therefore, it is possible to attract the vertical barnacle larvae existing in a wide range in the seawater to a desired position.

  According to the attracting base material for the vertical barnacle larvae according to claim 4, it is possible to induce the widespread growth of the vertical barnacle larvae in a wide range and attract them to the attracting base material. Therefore, it is possible to attract the vertical barnacle larvae existing in a wide range in the seawater to a desired position.

  According to the method for attracting a vertical barnacle larvae according to claim 5, since a concentration gradient of the protein centering on the attracting target region is formed, the epithelialization of the vertical barnacle larvae is induced extensively, Can be attracted to the area. Therefore, it is possible to attract the longhorned barnacle larvae existing in a wide range in the seawater to a desired attracting target area.

  The best mode for carrying out the present invention will be described below in detail with reference to the drawings.

  The protein of the present invention is a protein obtained from an extract of an adult vertical barnacle, having the amino acid sequence of SEQ ID NO: 1 on the N-terminal side and having a molecular weight of 32 kDa.

  There is no protein on the EMBL / Genbank / DDBJ protein database that matches or is similar to the protein having the amino acid sequence of SEQ ID NO: 1 on the N-terminal side. Therefore, the protein of the present invention is a novel protein.

  The molecular weight of the protein of the present invention is 32 kDa, which is different from that of the sugar chain-binding protein complex subunits (proteins with molecular weights of 76, 88 and 98 kDa) disclosed in Non-Patent Document 2. ing. The molecular weight of 32 kDa means a molecular weight measured by SDS-polyacrylamide gel electrophoresis.

  The protein of the present invention can be obtained by subjecting an adult extract of a vertical barnacle to a protein concentration, followed by protein purification using column chromatography.

  The adult Tatejima barnacle adult extract is obtained by homogenizing the entire tissue of the adult Tatejima barnacle in Tris-HCl buffer and then collecting the supernatant by centrifugation. A plurality of proteins including the protein of the present invention are mixed in the adult extract of the vertical barnacles.

  Protein concentration can be performed, for example, by salting out using ammonium sulfate.

Column chromatography is performed in the order of gel filtration, anion exchange chromatography, and hydroxyapatite column chromatography. A NaCl solution may be used as an eluent for anion exchange chromatography. Further, a KH ₂ PO ₄ solution may be used as an eluent for hydroxyapatite column chromatography.

  As gel particles used for gel filtration, for example, TOYOPEARL HW-55F manufactured by Tosoh Corporation can be used, but is not limited thereto, and has the same or similar properties and functions as the gel particles. Can be used.

  As the column used for anion exchange chromatography, for example, TOYOPEARL SuperQ-650M manufactured by Tosoh Corporation can be used, but is not limited to this, and has the same or similar properties and functions as this column. Can be used.

  As a column used for hydroxyapatite column chromatography, for example, Macro-prep ceramic hydroxyapatite, Type II, 40 μm manufactured by Bio-Rad Laboratories, Inc. can be used, but is not limited thereto, and is the same as this column. Alternatively, those having similar properties and functions can be used.

  1-3 show the protein elution pattern from each column chromatography and the fraction containing the protein of the present invention (shaded area in the figure). By a column chromatography operation, a protein having the amino acid sequence of SEQ ID NO: 1 on the N-terminal side and having a molecular weight of 32 kDa can be isolated.

  The protein of the present invention is a diffusive epithelial-inducing pheromone-like protein that exhibits the function of inducing the establishment of vertebrate barnacle larvae when dissolved and dispersed in seawater. Therefore, unlike the sugar chain-binding protein complex SIPC disclosed in Non-Patent Document 2, it is not necessary to adsorb to a substrate or the like, and the protein is diffused over a wide area in seawater by being dispersed in seawater. It is possible to induce a wide range of larvae of the vertical barnacle larvae.

  Here, although the density | concentration of the protein of this invention which induces the growth of a vertical barnacle larvae is at least 0.1 microgram / mL, it is preferable to set it as 1.0 microgram / mL or more. When the concentration is less than 0.1 μg / mL, it is not possible to sufficiently induce the establishment of the vertical barnacle larvae. By setting the concentration to 1.0 μg / mL or more, it is possible to induce about 70% or more of the vertical barnacle larvae. The upper limit of the protein concentration is not particularly limited, but it has been confirmed that about 80% of the vertical barnacle larvae can be induced at 10 μg / mL, and 1.0 μg / mL to 10 μg / mL. By doing so, it is possible to sufficiently induce the growth of the vertical barnacle larvae, which is preferable.

  In addition, by continuously supplying the protein of the present invention to the attracting target area of the vertical barnacle larvae and forming a protein concentration gradient centering on the attracting target area, it is possible to attract the vertical barnacle larvae to the attracting target area. it can.

  By continuously supplying the protein of the present invention to the attracting target area of the vertical barnacle larvae, the protein diffuses into the seawater from the attracting target area, and a protein concentration gradient centering on the attracting target area is formed. That is, a concentration gradient in which the protein concentration decreases as the attracting target region is set to the high concentration side and the distance from the attracting target region is formed. Therefore, it is possible to induce widespread growth of the vertical barnacle larvae by the protein diffused over a wide range in the seawater, and attract it to the attracting target region on the high concentration side of the protein concentration gradient. By utilizing this action, the vertical barnacle larvae can be attracted to a desired position from a wide range in seawater. The attracted vertical barnacle larvae can be captured by arranging a base or netting member on which the vertical barnacle larvae can grow in the attracting target area.

  Examples of a method for continuously supplying the protein of the present invention to the target area of the vertical barnacle larvae include, for example, a method of feeding an aqueous solution of the protein of the present invention to the target area of the attractant using a liquid feed pump, Examples include a method in which an aqueous solution of the protein of the present invention is accommodated in an airtight container equipped with an ultrafiltration membrane having a pore size capable of permeating protein, and the protein is gradually passed through the ultrafiltration membrane and supplied.

  Alternatively, the protein of the present invention may be supported on a carrier to serve as an attractant, and this attractant may be disposed in the attracting target region.

  As the carrier, a carrier that can carry the protein of the present invention, does not inhibit the elution of the protein of the present invention into seawater, and does not inhibit the induction of the vertical larvae larvae can be used. . For example, natural polymers such as agarose, collagen, fibrin, albumin, casein, cellulose fiber, cellulose triacetate, agar, calcium alginate, carrageenan, polyacrylamide, poly-2-hydroxyethyl methacrylic acid, polyvinyl chloride, γ- Examples include, but are not limited to, synthetic polymers such as methylpolyglutamic acid, polystyrene, polyvinylpyrrolidone, polydimethylacrylamide, polyurethane, and photocurable resins (polyvinyl alcohol derivatives, polyethylene glycol derivatives, polypropylene glycol derivatives, polybutadiene derivatives, etc.). It is not something.

  Alternatively, the protein of the present invention may be formed into a solid form to serve as an attractant, and this attractant may be disposed in the attracting target area.

  The solid attractant can be obtained, for example, by mixing the protein of the present invention with a binder material or the like and molding it by pressure, but the method for producing the solid attractant is not limited to this method. Absent.

  Alternatively, the protein of the present invention may be attached to the surface of a base material to form an attracting base material, and this attracting base material may be arranged in the attracting target region. In this case, if the material of the base material is a material capable of growing the vertical barnacle larvae, it is possible to capture the vertical barnacle larvae on the attracting base material itself without disposing a growing member in the attracting target area.

  The substrate has a certain degree of adsorptivity and elution for the protein of the present invention, that is, the protein of the present invention is held on the surface for a certain period of time and placed on the attracting target area. A base material from which the protein of the present invention is gradually eluted may be used. Specific examples include concrete, slate, and the like, but are not limited thereto.

  In this way, the habitat of the vertical barnacle larvae can be changed by attracting the vertical barnacle larvae to the desired attracting target area, and the marine structure, the cooling water system of the thermal power / nuclear power plant, ship, fish net, etc. It is possible to suppress the attachment of larvae. The present invention can also be used for the purpose of positively capturing the vertical barnacles. In this case, for example, by attaching the vertical barnacle larvae to the base material and dissolving only the base material, it is possible to save the trouble of peeling the vertical barnacle larvae from the base material. Alternatively, the vertical barnacle larvae may be attached to a substrate and grown to an adult, and then the substrate may be dissolved to obtain an adult. In other words, by using the present invention to attach the vertical barnacle auricle to the substrate, the vertical barnacle can be obtained at a desired growth stage. This facilitates the industrial demands of the vertical barnacles, for example, the supply of raw materials such as feed to cultured seafood.

  The above-described embodiment is an example of a preferred embodiment of the present invention, but is not limited thereto, and various modifications can be made without departing from the scope of the present invention. For example, a sugar chain-binding protein complex composed of 76, 88 and 99 kDa subunits that induces the growth of vertical barnacle larvae by adsorbing to the substrate or the like on the surface of the substrate for establishment placed in the attraction target region You may make it adsorb | suck body SIPC (refer nonpatent literature 2). In this case, the vertical barnacle larvae attracted to the attracting target area can be further attracted by the substrate for establishment, and the vertical barnacle larvae can be attracted and captured effectively.

  Moreover, you may make it utilize the attracting method of the vertical barnacle larva of this invention for the prediction technique of an adhesion time. For example, by using the method for attracting larvae larvae of the present invention to monitor the change over time of the population variation of larvae larvae attracted in a specific sea area, the change over time of the population change and the time of attachment to the marine construct, etc. By accumulating data indicating the relationship, the adhesion time can be predicted.

  In addition, since the protein of the present invention has a function of specifically inducing the establishment of the vertical barnacle larvae, the species of barnacle larvae that are extremely difficult to perform species determination based only on morphological taxonomic differences It can also be used for determination technology.

  EXAMPLES The present invention will be described below with reference to examples, but the present invention is not limited to these examples.

(1. Protein purification)
After 1.5-fold (w / v) Tris-HCl buffer solution (50 mM, pH 7.5) was added to the whole tissue of the adult vertical barnacle, the supernatant was collected by centrifugation. Next, ammonium sulfate solution (70% by weight) was added to the recovered supernatant for salting out, and the recovered supernatant protein was concentrated.

Next, the protein was purified using column chromatography to obtain a protein having a molecular weight of 32 kDa. Column chromatography was performed in the order of (A) to (C) shown below.
(A) Gel filtration (B) Anion exchange chromatography (C) Hydroxyapatite column chromatography

  Gel filtration was performed using TOYOPEARL HW-55F manufactured by Tosoh Corporation as gel particles. The protein elution pattern in gel filtration is shown in FIG. The protein eluate in the hatched area (33 mL to 63 mL) was collected as a fraction.

  Anion exchange chromatography was performed using TOYOPEARL SuperQ-650M manufactured by Tosoh Corporation as a column and a NaCl solution as an eluent. The protein elution pattern in anion exchange chromatography is shown in FIG. The protein eluate in the shaded area (160 mL to 185 mL) was collected as a fraction. In addition, the broken line in FIG. 2 represents the gradient of the concentration (right vertical axis, unit mM) of the NaCl solution used as the eluent.

Hydroxyapatite column chromatography, Bio-Rad Laboratories Ltd. of Macro-prep ceramic hydroxy apatite, TypeII , a 40μm used as a column was performed using _KH 2 PO ₄ solution as eluent. The protein elution pattern in hydroxyapatite column chromatography is shown in FIG. The protein eluate in the region indicated by oblique lines (120 mL to 145 mL) was collected as a fraction. In addition, the broken line in FIG. 3 represents the gradient of the concentration (right vertical axis, unit mM) of the KH ₂ PO ₄ solution used as the eluent.

  FIG. 4 shows the results of measuring the molecular weight of the protein purified by the column chromatography by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). In FIG. 4, the electrophoresis result on the left shows the molecular weight marker, and the electrophoresis result on the right shows the protein purified by the column chromatography. From this measurement result, it was confirmed that the molecular weight of the purified protein was about 32 kDa. In addition, other molecular weight proteins were not detected. From this, it was clarified that the protein of 32 kDa can be purified with high purity by using the column chromatography.

(2. Confirmation of induction-inducing activity)
A test was carried out to confirm the epithelial-inducing activity of the 32 kDa protein purified by the above method (hereinafter sometimes simply referred to as a purified protein) against the vertical barnacle larvae.

Prepare a 24-hole plate made of polystyrene, and store 1 mL of sterilized seawater under the following conditions (A) in 6 of the 24 holes, and sterilized seawater under the following conditions (B) in another 6 holes 1 mL each was accommodated, and 1 mL of sterile seawater having the following condition (C) was further accommodated in another 6 holes, and 1 mL of sterile seawater having the following condition (D) was accommodated in the remaining 6 holes.
(A) Sterile seawater not containing purified protein (B) Sterile seawater with a purified protein concentration of 0.1 μg / mL (C) Sterile seawater with a purified protein concentration of 1.0 μg / mL (D) Purified protein concentration of 10 μg / ML sterile seawater

In each hole, 7 to 10 cypris larvae of vertical barnacles were housed and allowed to stand at 25 ° C. in a light-shielded state. The state of cypris larvae after 3 hours was observed with a stereomicroscope, and the number of individuals attached to the plate wall of each hole was counted. The adhesion rate was calculated by Equation 1 shown below.
[Formula 1] Adhesion rate (%) = number of attached individuals / total number of individuals × 100

  This test was performed 3 times, and the average value of the adhesion rate was obtained.

  The test results are shown in FIG. FIG. 5 shows the adherence rate of Cypris larvae of the vertical barnacles to sterilized seawater under the above conditions (A) to (D), and the higher the adherence rate, the higher the purified protein has higher induction activity of Cypris larvae. Means. From this result, when using sterilized seawater with a purified protein concentration of 0.1 μg / mL, the adhesion rate was slightly higher than when using sterilized seawater not containing purified protein. It was clarified that the use of sterilized seawater having a purified protein concentration of 1.0 μg / mL improved the adhesion rate by a factor of 3 or more as compared with the case of using sterilized seawater containing no purified protein. Furthermore, it was revealed that the use of sterilized seawater having a purified protein concentration of 10 μg / mL improved the adhesion rate by about 4 times compared to the case of using sterilized seawater containing no purified protein.

  From the above results, it was clarified that the purified protein obtained in this example exhibits an induction-inducing activity when dissolved in seawater against Cypris larvae, which are the juvenile larvae of the vertical barnacles.

  And it became clear that the effect of inducing the establishment of Cypris larvae can be obtained by setting the purified protein concentration of seawater to 0.1 μg / mL or more. It was revealed that this effect was greatly improved by setting the purified protein concentration of seawater to 1.0 μg / mL and further improved by setting it to 10 μg / mL.

  Moreover, when the same experiment was conducted after heating the purified protein at 90 ° C. for 10 minutes, it was confirmed that the same result as that obtained when the purified protein was not heated was obtained. From this, it was clarified that this protein does not cause inactivation (protein denaturation) by heat treatment, and hardly inactivates due to the use environment temperature. Therefore, it was found that the protein can be used without worrying about the temperature of the environment in which the protein is actually used. Moreover, it was suggested that the protein could be purified with high purity by simplifying the purification process of the protein using the heat resistance of the protein, that is, by heat-inactivating other proteins.

(3. Confirmation of attracting effect by purified protein)
The effect of the purified protein on the attraction of the vertical barnacle larvae was examined.

  A conceptual diagram of the test method is shown in FIG. 100 mL of sterilized seawater (symbol b) and 43 Cypris larvae of vertical barnacles were housed in a glass petri dish (symbol a) having a diameter of 12 cm. Then, two agarose gels (1) with 1 mL of 1% agarose formed into a cylindrical shape (reference symbol c) and one purified protein-containing agarose gel with 10 μg of purified protein mixed with 1 mL of 1% agarose (1) The symbol d) was prepared, and these agarose gels were arranged at regular intervals slightly inside the circumference of the glass petri dish.

  The glass petri dish was allowed to stand at 25 ° C. for 24 hours in a light-shielded state, and then the number of Cypris larvae adhering to the glass petri dish wall was counted. The results are shown in FIG. In FIG. 7, other than agarose gels (symbols c and d) indicate positions where Cypris larvae are attached to the glass petri dish wall.

  Cypris larvae adhering to the glass petri dish were 27 out of 43 individuals. Further, it was confirmed that 85% of the adhered individuals adhered to a region in the vicinity where the purified protein-containing agarose gel d was disposed.

  From the purified protein-containing agarose gel d, the purified protein is gradually eluted and diffused over a wide range in the sterilized seawater, and a protein concentration gradient centering on this gel is formed. That is, in this concentration gradient, the protein concentration decreases with increasing distance from the gel, with the periphery of the gel being on the high concentration side. In this example, 85% of the adherent individuals were attached to the area in the vicinity where the gel was placed, so that the induction of Cypris larvae was induced extensively by the action of the purified protein diffused over a wide range. It was clarified that the vertical barnacle larvae can be attracted to the high concentration region of the protein concentration gradient.

(4. Identification of amino acid sequence)
For the purified protein obtained in this example, the amino acid sequence of the N-terminal portion was analyzed. The amino acid sequence was analyzed by Edman reaction using a protein sequencer PPSQ21 manufactured by Shimadzu Corporation after reducing the pyridylethylation of the purified protein.

As a result, it was confirmed that the amino acid sequence of 33 residues on the N-terminal side was the amino acid sequence represented by SEQ ID NO: 1 in the sequence listing. When a single letter abbreviation is used, this amino acid sequence is represented as follows.
(H) -EPQPPGVPSTPQLAGVLTNLYRRFLTVTAPKQLN

  When this amino acid sequence was searched on the EMBL / Genbank / DDBJ protein database, there was no matching or similar sequence. From this, it was considered that the purified protein obtained in this example was a novel protein whose sequence was unknown.

  Based on the above results, the purified protein obtained in this example, that is, the protein obtained from the adult extract of the vertical barnacle, having the amino acid sequence of SEQ ID NO: 1 on the N-terminal side and having a molecular weight of 32 kDa Was thought to be a novel protein of unknown sequence.

It is a figure which shows the protein elution pattern in gel filtration. It is a figure which shows the protein elution pattern in anion exchange chromatography. It is a figure which shows the protein elution pattern in a hydroxyapatite column chromatography. It is a figure which shows the result at the time of using for the protein of this invention for SDS-PAGE. It is a figure which shows the result of an epidemic induction activity confirmation test. It is a figure which shows the concept of the method of an attraction effect confirmation test. It is a figure which shows the result of an attraction effect confirmation test.

Claims (5)

  A protein obtained from an extract of an adult tateji barnacle and having the amino acid sequence of SEQ ID NO: 1 on the N-terminal side and having a molecular weight of 32 kDa.   An attractant for vertebral larvae larvae, wherein the protein according to claim 1 is supported on a carrier.   The attractant for the vertical barnacle larvae which the protein of Claim 1 was shape | molded in solid form.   An attracting base material for vertebrate barnacle larvae, wherein the protein according to claim 1 is attached to the surface of the base material.   A method for attracting vertebrate barnacle larvae, comprising continuously supplying the protein according to claim 1 to an attraction target area of a vertical barnacle larva to form a concentration gradient of the protein centering on the attraction target area.