近江牡蛎i-型溶菌酶基因分析及温度对基因表达的影响

doi:10.11978/j.issn.1009-5470.2012.06.011

Abstract

Abstract: Lysozyme is one of the important effectors involved in a variety of immune responses in innate immune system of organisms. A conservative i-type lysozyme cDNA sequence was cloned from Crassostrea hongkongensis and its expression in different tissues was studied. The effects of different temperatures on the gene expression in the experiment were also studied. The cloned conservative i-type lysozyme cDNA sequence contained 634 bp with a 420 bp open reading frame and the cDNA encoded 139 amino acids. A new conservative amino acid sequence, HNGGPRGC, and the typical features were found in the protein sequence of the i-type lysozyme from C. hongkongensis, including specific amino acid residue sequence, CL(E/L/R/H)C(I/M)C, a partial highly conservative sequence, SCG(P/Y)FQI, and the enzyme active site (Glu³⁴, Asp⁴⁵, Ser⁴⁸, Trp⁶¹). Real-time quantitative PCR was used to assess the expression pattern of the i-type lysozyme gene in the digestive gland, muscle, gill, mantle, and labial palps of C. hongkongensis. The highest expression level of the i-type lysozyme gene was found in the digestive gland, the second in the gill, and the lowest in the muscle. A temperature stress experiment was conducted to study the expression control of the i-type lysozyme gene. The expression level was stable when the water temperature was at 20?25℃. A tendency to decrease expression was found at 6 or 13℃. The expression was maintained at a relatively high level throughout of 96 h at 27℃. The expression level was found the highest at 12 h and sharply declined after 48 h at 34℃. It is concluded that the expression of the i-type lysozyme gene was affected by temperature, which could be used to control the immune function of C. hongkongensis in response to external environment

Key words: i-type lysozyme, clone, expression, temperature, Crassostrea hongkongensis

CLC Number:

P735

XU You-qing, WU Wei-jun, DING Zhao-kun, PAN Zhi-zhong, HE Wei, WU Tie-jun, CHEN Xiu-li, LI Yong-mei, JIANG Wei-ming. Analysis of i-type lysozyme gene from Crassostrea hongkongensis and the effect of temperature on the gene expression[J].Journal of Tropical Oceanography, 2012, 31(6): 69-75.

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Analysis of i-type lysozyme gene from Crassostrea hongkongensis and the effect of temperature on the gene expression

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