Molecular cloning and expression profile of putative antilipopolysaccharide factor in Chinese shrimp(Fenneropenaeus chinensis).
A new antimicrobial protein gene of the anti-lipopolysaccharide factor family (tentatively named as ALFFc) has been cloned from hemocytes of the Chinese shrimp Fenneropenaeus chinensis by rapid amplification of 3' and 5' complementary DNA ends with polymerase chain reaction. The full-length complementary DNA of ALFFc consists of 600 bp with a 369-bp open reading frame, encoding 123 amino acids. The deduced peptide contains a putative signal peptide of 25 amino acids and mature peptide of 98 amino acids. The molecular mass of the deduced mature peptide is 13799.16 Da. It is highly cationic, with a theoretical pI of 10.3. The deduced amino acid sequence of ALFFc showed 56% homology with sequences of Tachypleus tridentatus and L. polyhemus. The tissue expression profile of this gene was studied by Northern blot, and ALFFc transcripts were mainly detected in hemocytes, gill, and intestine. RNA in situ hybridization showed that ALFFc was constitutively expressed in hemocytes. Capillary electrophoresis reverse transcriptase PCR was used to quantify the variation of messenger RNA transcription level during the artificial infection process with Vibrio anguillarum. Significant enhancement of ALFFc transcription appeared during the first 24 hours in response to Vibrio infection. These results provide useful information for understanding the function of ALFFc in shrimp.
Liu
Fengsong
F
Experimental Marine Biology Laboratory, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.
Liu
Yichen
Y
Li
Fuhua
F
Dong
Bo
B
Xiang
Jianhai
J
D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides; D060829:Arthropod Proteins; D001483:Base Sequence; D015152:Blotting, Northern; D002681:China; D003001:Cloning, Molecular; D017931:DNA Primers; D020869:Gene Expression Profiling; D017403:In Situ Hybridization; D007447:Invertebrate Hormones / Q000235:genetics*; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
D023181:Antimicrobial Cationic Peptides; D060829:Arthropod Proteins; D017931:DNA Primers; D007447:Invertebrate Hormones; C034705:antilipopolysaccharide factor (Limulus)