论文

革胡子鲶生长激素全长cDNA 的克隆与序列分析

  • 张茂群 ,
  • 陈承勋 ,
  • 郭永军 ,
  • 郭菁 ,
  • 王晓梅
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  • 天津农学院 水产科学系,天津市水产生态及养殖重点实验室,天津 300384
张茂群(1984-),男,山东文登人,在读硕士,主要从事水产动物分子生物学的研究.

收稿日期: 2009-09-29

  网络出版日期: 2014-10-14

基金资助

天津市应用基础及前沿技术研究计划重点研究项目(08JCZDJC19000)

Cloning and Sequence Analysis of Full-length Growth Hormone cDNA from Clarias gariepinus

  • ZHANG Mao-qun ,
  • CHEN Cheng-xun ,
  • GUO Yong-jun ,
  • GUO Jing ,
  • WANG Xiao-mei
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  • Department of Fisheries Science, Tianjin Agricultural University, Tianjin Key Laboratory of Aqua-Ecology and Aquaculture, Tianjin 300384, China

Received date: 2009-09-29

  Online published: 2014-10-14

摘要

通过RT-PCR和RACE-PCR的方法,克隆了革胡子鲶生长激素基因全长cDNA,其长度为973 bp,包含一个03 bp的开放阅读框架(Open reading frame,ORF),59 bp的5′非编码区和311 bp的3′非编码区(含PolyA 尾25 bp).将革胡子鲶GH cDNA的ORF、5′非编码区和3′非编码区的序列分别与同为鲶形目印度囊鳃鲶、巨鲶鱼、南方鲶和鲶的上述序列进行比对分析,结果表明:革胡子鲶与上述鱼类生长激素ORF的核苷酸与氨基酸序列同源性较高,平均值分别为91.2%和9.4%,ORF区核苷酸的碱基替代类型表现出T/C转换的偏向性,平均值为44.5%,同时表现出转换/颠换偏差,平均值为2.381.5′和3′非编码区序列同源性平均值分别为75.4%和77.0%,保守性低于编码区.

本文引用格式

张茂群 , 陈承勋 , 郭永军 , 郭菁 , 王晓梅 . 革胡子鲶生长激素全长cDNA 的克隆与序列分析[J]. 华北农学报, 2009 , 24(6) : 27 -32 . DOI: 10.7668/hbnxb.2009.06.006

Abstract

In this study,the GHfull2length cDNA is cloned with the employment of RT2PCR and RACE2PCR techniques,which is 973 bp in length.The total length cDNA includes an open readingframe (ORF) of 603 bp,5′ UTRof 59 bp and 3′ UTR of 311 bp (Poly A of 25 bp included).Pairwise and multiple alignments of the sequences of ORF,5′ UTR,3′ UTR of Clarias gariepinus GH cDNA with those of Heteropneustes fossilis,Pangasianodon gigas,Silurus meridionalis and Silurus asotus are performed using DNAMAN 5.2.and MEG A 411 software.Sequence alignment analysis indicate that the average homologies are 91.% and 9614 % separately for ORF nucleotide and amino acid sequence between Clarias gariepinus and the 4 species of Siluriforms catfish mentioned above.ORF nucleotide substitution patterns represent the bias of transition between T and C and the average value is 44.5 %.Meanwhile,the base substitutions also show transition/ transversion bias and the average ratio reaches 2.381.5′ UTR and 3′ UTR nucleotide sequence homology are 7514 % and 7710 %,respectively,which are lower conservative than that of ORF nucleotide sequence.

参考文献

[1] 李恒国,朱兴国,闵宪伟.高密度集约化革胡子鲶养殖技术[J].渔业致富指南,2006(8):31 -32.
[2] 韦家永,薛良义.鱼类生长激素的研究概况[J].浙江海洋学院学报:自然科学版,2004(3):56-59.
[3] 王树启,许友卿,丁兆坤.生长激素对鱼类的影响及其在水产养殖中的应用[J].水产科学,2005,24(7):42-46.
[4] Sekine S,Mizukami T,Nishi T,et al.Cloning and expression of cDNA for salmon growth hormone in Escherichia coli[J].Proc Natl Acad Sci USA,1985,82:4306-4310.
[5] Agellon L B,Davies S L,Lin C M,et al.Rainbow trout has two genes for growth hormone[J].Mol Rep Develop,1988,1:11-17.
[6] 马细兰,张勇,黄卫,等.尼罗罗非鱼生长激素及其受体的cDNA 克隆与mRNA 表达的雌雄差异[J].动物学报,2006,52(5):924-933.
[7] May D,Alrubaian J,Patel S,et al.Studies on the GH/SL gene family:cloning of African lungfish(Protopterus annectens)growth hormone and somatolactin and Toad(Bufo marinus)growth hormone[J].Gen Comp Endocrinol,1999,113:121-135.
[8] Ayson F G,de Jesus E G T,Amemiya Y,et al.Isolation,cDNA cloning,and growth promoting activity of rabbitfish (Siganus guttatus) growth hormone[J].Gen Comp Endocrinol,2000,117:251-259.
[9] Tang Y,Lin C M,Chen T T,et al.Structure of the channel catfish (Ictalurus punctatus) growth hormone gene and its evolutionary implications[J].Mol Marine Biol Biotechnol,1993,2(4):198-206.
[10] Lemaire C,Warit S,Panyim S.Giant catfish Pangasianodo gigas growth hormone-encoding cDNA:cloning and sequencing by one-side polymerase chain reaction[J].Gene,1994,149(2):271-276.
[11] 张为民,张勇,李欣,等.斜带石斑鱼生长激素cDNA 克隆及其在大肠杆菌的融合表达[J].水产学报,2003,27(5):392-396.
[12] 杜启艳,陈丽丽,南平,等.鲶生长激素基因cDNA 的克隆和原核表达[J].水产科学,2007,26(4):195-199.
[13] Johansen B,Johnsen O C,Valla S.The complete nucleotide sequence of the growth-hormone gene from Atlantic salmon (Salmo salar)[J].Gene,1989,77:317-324.
[14] Chao S C,Pan F M,Cheng W C.Purification of carp growth hormone and cloning of the complementary DNA[J].Biochim Biophys Act,1989,1007(2):233-236.
[15] Zhang J N,Song P,Hu J R,et al.Molecular cloning and sequene analysis of full-length growth hormone cDNAs from six important economic fishes[J].Acta Genetica Sinica,January 2005,32(1):19-29.
[16] 高春生,范光丽,杨国宇,等.淇河鲫生长激素(growth hormone)全长cDNA 的克隆与序列分析[J].分子科学学报,2008,24(1):21-26.
[17] 宋平,胡隐昌,向筑,等.南方鲶生长激素完整cDNA 的克隆及其DNA 序列的分析[J].水生生物学报,2002,26(3):272-279.
[18] 杨学明,张立,黄光华,等.革胡子鲶生长激素基因克隆与序列同源分析[J].西南农业学报,2008,21(2):483-486.
[19] Kao F T,Yu J W.Chromosome microdissection and cloning in human genome and genetic disease analysis[J].Proc Nati Acad Sci USA,2005,88(3):1844-1848.
[20] 杨学明,何荆洲,黄雄军,等.革胡子鲶生长激素cDNA 克隆与蛋白质结构分析[J].遗传,2008,30(7):913-918.
[21] 唐萍,王强,陈建群.茄科植物叶绿体基因组插入、缺失和核苷酸替代的发生方式及影响[J].遗传,2008,30(11):1506-1512.
[22] Collins D W,Jukes T H.Rates of transition and trans,version in coding sequences since the human-rodent divergence[J].Genomics,1994,20(3):386-396.
[23] 刘福平,白俊杰.单核苷酸多态性及其在水产动物遗传育种中的应用[J].中国水产科学,2008,15(4):704-712.
[24] 王伟,汪亚平,朱作言.鱼类生长激素基因工程研究[J].自然科学进展,2002,12(5):456-460.
[25] 马飞,武耀廷,徐晓风.遗传密码子和氨基酸若干物理化学特性的相关性研究[J].安徽农业大学学报,2003,30(4):439-445.
[26] 刘建文,施用晖,乐国伟.动物生长轴的激素调控[J].中国饲料,2003(14):7-9.
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