Recombinant perlucin nucleates the growth of calcium carbonate crystals: molecular cloning and characterization of perlucin from disk abalone, Haliotis discus discus.
Perlucin is well known as an important functional protein regulating pearl formation and shell biomineralization. In this study, we cloned the perlucin gene from the abalone Haliotis discus discus cDNA library. The full-length cDNA of the abalone H. discus discus perlucin gene consisted of 1038 bp nucleotides, encoding a putative signal peptide of 22 amino acids and a mature protein of 129 amino acids, which shared 55% identity with the homologous protein in greenlip abalone. The mature protein coding sequence was inserted into pMal-c2X expression vector and it expressed the recombinant protein in E. coli (Rosetta-gammi DE3). The maltose binding protein (MBP) fusion perlucin successfully promoted calcium carbonate precipitation and directed calcite crystal morphological modification. The successful expression of active recombinant perlucin suggested that recombinant perlucin gene transfer has the capability by color modification to improve the pearl's value. In the view of molecular structure, perlucin was a typical C-type lectin, which contained one highly conserved carbohydrate recognition domain. Reverse transcription polymerase chain reaction (RT-PCR) results showed that perlucin gene was expressed not only in the mantle, but also in the gill and digestive tract. Further characterization of perlucin in abalone non-self recognition and disease resistance is promising and anticipated.
Wang
Ning
N
Department of Aquatic Life Medicine, College of Ocean Science, Cheju National University, Jeju-si 690-756, Republic of Korea.
Lee
Youn-Ho
YH
Lee
Jehee
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002119:Calcium Carbonate / Q000737:chemistry*; D011232:Chemical Precipitation; D003001:Cloning, Molecular; D003460:Crystallization; D049851:Gastropoda / Q000737:chemistry / Q000235:genetics*; D037102:Lectins / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D037102:Lectins; D011994:Recombinant Proteins; C403488:perlucin, Haliotis laevigata; D002119:Calcium Carbonate