Inaugural ASAS–CAAV Asia Pacif ic Rim Conference Abstracts

T291 CBHI gene cloning and sequence analysis ofPenicillium oxalicum Currie et Thom. B. W. Wang*, Q. Zhang, B. Yue, W.H. Ge, and M. A. Zhang, High Quality Waterfowl Research Institute, QingdaoAgricultural University, Qingdao, Shandong Province, China.Exoglucanase is an important tool for fungi to degrade natural cellulose. Tostudy the molecular character of exoglucanase, Penicillium oxalicum Currie etThom F67 from geese was used to test strains. Degenerating PCR was used toamplify the gene fragment of CBHI whose sequence length was as long as 1059bp. On the basis of the CBHI gene fragment, the cDNA of Penicillium oxalicumCurrie et Thom F67 was used as a template to clone the 3′ and 5′ flankingsequences through thermal asymmetric interlaced PCR. The sequence analysisshowed that the length of 3′ and 5′ flanking sequences were 602 bp and 728 bp,Poultry Genetics Postersrespectively. In addition, the full-length CBHI was cloned by through reversetranscription-PCR (EU727171). Sequence analysis showed that the full lengthof this sequence was as long as 1638 bp and that the gene encoded 545 aminoacids and a stop codon, TAA. It had the highest homology with the Penicilliumjanthinellum gene, which could reach 80%. The result of the full-length CBHIsequence enriched the bioinformatics; the building of a eukaryotic expressionvector laid a foundation for further investigation of the expression systems andobtaining an efficient engineering strain.Key Words: goose origin Penicillium oxalicum Currie et Thom, exoglucanase,thermal asymmetric interlaced PCRT292 Effect of vitamin D 3by injection on the β defensins inTaihe Silky fowl. S. Li* 1 , L. Ouyang 1 , D. Zhou 2 , J. Xie 1 , and Q. Wei 1 , 1 Instituteof Animal Husbandry and Veterinary, Jiangxi Academy of Agricultural Science,Nanchang City, Jiangxi, China, 2 College of Animal Science and Technology,Sichuan Agricultural University, Yaan City, Sichuan, China.The expression of avian β defensins (AVBD), Toll-like receptors (TLR), andvitamin D receptor (VDR) was studied following in vivo with injection vitaminD 3. The AVBD are important effector molecules of innate immunity and playa critical role in the host against invasion of pathogenic microorganisms.Vitamin D 3as an immunomodulator could directly induce the expression ofantimicrobial peptide and related immune factors in mammalian monocytesor epithelial cells in poultry, lacking the 1-α-hydroxylase of vitamin D 3intothe 1,25 dihydroxy-vitamin D 3. Healthy 42-d-old Silky fowl were abdominallyinjected with vitamin D 3or were untreated. Real-time PCR analyses revealedthat injection of vitamin D 3significantly up-regulated the expression (P < 0.05)of TLR ( TLR2, TLR6, and TLR7),VDR, AVBD (AVBD 1, AVBD 5, AVBDPoultry Immunology Posters7, AVBD 9, AVBD 10, and AVBD 12), and 24-hydroxylase (CYP24A1) inthe gut (duodenum, jejunum, ileum, and cecum) during different times from8 to 24 h postinjection, similar to the expression of TLR (TLR2, TLR5),VDR,AVBD 6, and 24-hydroxylase (CYP24A1) in the liver, spleen, and kidney.These results suggest that expression of VDR, AVBD, and TLR seems to beinduced by vitamin D 3, and we concluded that the tissues expressing TLR andVDR respond to vitamin D 3and, in turn, up-regulate tissue cellular functionsto synthesize AVBD. Intraperitoneal injection of vitamin D 3likely resulted inenhanced expression of AVBD, TLR, and VDR, which provided insight intofactors important to the control of the innate immune response in the chicken.This work was supported by Support Plan Fund of Jiangxi and Innovation Fundfrom Jiangxi Academy of Agricultural Sciences, China.Key Words: induction, chicken, avian betadefensins84