一种栉孔扇贝丝氨酸蛋白酶基因的染色体定位及其内部SNP的研究
收稿日期: 2009-03-06
修回日期: 2009-10-09
网络出版日期: 2009-11-17
基金资助
国家自然科学基金重点项目(编号:30730071)和国家高技术研究发展计划(863)项目(编号:2007AA09Z430)资助
Chromosomal localization and development of SNP markers of a serine protease gene in Farrer’s scallop (Chlamys farreri)
Received date: 2009-03-06
Revised date: 2009-10-09
Online published: 2009-11-17
病害问题是制约我国扇贝养殖业发展的关键, 因此贝类的先天免疫也成为当前研究的热点。丝氨酸蛋白酶是先天免疫中至关重要的酶类,在许多通路中起信号放大作用。目前对栉孔扇贝丝氨酸蛋白酶的研究主要集中于基因序列分析和表达谱研究, 基因定位方面的研究尚未开展。文章以包含一种栉孔扇贝丝氨酸蛋白酶基因的BAC克隆为探针, 利用BAC-FISH技术将其定位到一对同源染色体的长臂上, 为丝氨酸蛋白酶基因的后续研究提供了基础; 同时, 利用PCR产物直接测序法筛选了该基因内部的6个SNP标记, 这些SNP标记可供遗传图谱定位使用, 从而可以实现遗传图谱与染色体间的锚定与初步整合
郇聘,张晓军,李富花,张洋,赵翠,刘保忠,相建海 . 一种栉孔扇贝丝氨酸蛋白酶基因的染色体定位及其内部SNP的研究[J]. 遗传, 2009 , 31(12) : 1241 -1247 . DOI: 10.3724/SP.J.1005.2009.01.004
Disease control is a main objective in the culture of Farrer’s scallop (Chalmys farreri). Many investigations have focused on the innate immune system of this species. Serine proteases amplify signals in many immune-related pathways and thus play a crucial role in innate immune system. Recently, studies on serine protease in C. farreri were mainly on sequence analysis and expression profile, but no study on gene localization was reported. In this study, a BAC clone (CFB040H03) that contains a serine protease gene of C. farreri was located on the long arms of a pair of homologous chromosomes by BAC-FISH technique. Six SNPs were found inside the serine protease gene using direct sequencing of the PCR products. The chromosomal localization of serine protease gene would provide fundamental support for further research on this gene. The SNPs obtained can be mapped to the genetic linkage map and is helpful to the construction of the integration map of C. farreri.
[1] 吴龙涛, 宋林生, 胥炜, 邱丽华, 李红蕾, 苏建国, 相建海. 栉孔扇贝热休克蛋白70基因cDNA的克隆与分析. 高技术通讯, 2003, 13(11): 75–79.
[2] Su JG, Song LS, Xu W, Wu LT, Li HL, Xiang JH. cDNA cloning and mRNA expression of the lipopolysaccharide- and beta-1,3-glucan-binding protein gene from scallop Chlamys farreri. Aquaculture, 2004, 239(1–4): 69–80.
[3] Gao Q, Song L, Ni D, Wu L, Zhang H, Chang Y. cDNA cloning and mRNA expression of heat shock protein 90 gene in the haemocytes of Zhikong scallop Chlamys far-reri. Comp Biochem Physiol B Biochem Mol Biol, 2007, 147(4): 704–715.
[4] Qiu L, Song L, Xu W, Ni D, Yu Y. Molecular cloning and expression of a Toll receptor gene homologue from Zhikong Scallop, Chlamys farreri. Fish Shellfish Immunol, 2007, 22(5): 451–466.
[5] Qiu L, Song L, Yu Y, Xu W, Ni D, Zhang Q. Identification and characterization of a myeloid differentiation factor 88 (MyD88) cDNA from Zhikong scallop Chlamys farreri. Fish Shellfish Immunol, 2007, 23(3): 614–623.
[6] Wang H, Song L, Li C, Zhao J, Zhang H, Ni D, Xu W. Cloning and characterization of a novel C-type lectin from Zhikong scallop Chlamys farreri. Mol Immunol, 2007, 44(5): 722–731.
[7] Yu Y, Qiu L, Song L, Zhao J, Ni D, Zhang Y, Xu W. Mo-lecular cloning and characterization of a putative lipopolysaccharide-induced TNF-a factor (LITAF) gene homologue from Zhikong scallop Chlamys farreri. Fish Shellfish Immunol, 2007, 23(2): 419–429.
[8] Zhang H, Song L, Li C, Zhao J, Wang H, Gao Q, Xu W. Molecular cloning and characterization of a thio-ester-containing protein from Zhikong scallop Chlamys farreri. Mol Immunol, 2007, 44(14): 3492–500.
[9] Zhang H, Song L, Li C, Zhao J, Wang H, Qiu L, Ni D, Zhang Y. A novel C1q-domain-containing protein from Zhikong scallop Chlamys farreri with lipopolysaccharide binding activity. Fish Shellfish Immunol, 2008, 25(3): 281–289.
[10] Liu CT, Hou RF, Ashida M, Chen CC. Effects of inhibitors of serine protease, phenoloxidase and dopa decarboxylase on the melanization of Dirofilaria immitis microfilariae with Armigeres subalbatus haemolymph in vitro. Parasi-tology, 1997, 115(Pt 1): 57–68.
[11] Levashina EA, Langley E, Green C, Gubb D, Ashburner M, Hoffmann J A, Reichhart JM. Constitutive activation of Toll-mediated antifungal defense in serpin-deficient Dro-sophila. Science, 1999, 285(5435): 1917–1919.
[12] Zhu L, Song LS, Zhao JM, Xu W, Chang YQ. Molecular cloning, characterization and expression of a serine prote-ase with clip-domain homologue from scallop Chlamys farreri. Fish Shellfish Immunol, 2007, 22(5): 556–566.
[13] Zhu L, Song LS, Mao YZ, ZhaoJM, Li CH, Xu W. A novel serine protease with clip domain from scallop Chlamys farreri. Mol Biol Rep, 2008, 35(2): 257–264.
[14] 王永平, 郭希明. FISH技术在贝类分子生物学研究中的应用. 生命科学研究, 2001, 5(4): 283–289.
[15] Sachidanandam R, Weissman D, Schmidt SC, Kakol JM, Stein LD, Marth G, Sherry, S, Mullikin, JC, Mortimore, BJ, Willey DL, Hunt SE, Cole CG, Coggill PC, Rice CM, Ning Z, Rogers J, Bentley DR, Kwok PY, Mardis ER, Yeh RT, Schultz B, Cook L, Davenport R, Dante M, Fulton L, Hillier L, Waterston RH, McPherson JD, Gilman B, Schaffner S, Van Etten WJ, Reich D, Higgins J, Daly MJ, Blumenstiel B, Baldwin J, Stange-Thomann N, Zody MC, Linton L, Lander ES, Altshuler D, International SNP Map Working Group. A map of human genome sequence varia-tion containing 1.42 million single nucleotide polymor-phisms. Nature, 2001, 409(6822): 928–933.
[16] Zhang Y, Zhang X, Scheuring CF, Zhang HB, Huan P, Li F, Xiang J. Construction and characterization of two bacte-rial artificial chromosome libraries of Zhikong s
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