鸡二价金属转运蛋白1 cDNA的克隆与序列分析

白世平;吕林;罗绪刚;刘彬

中国生物化学与分子生物学报 ›› 2008, Vol. 24 ›› Issue (06) : 574-580.

PDF(517 KB)
PDF(517 KB)
中国生物化学与分子生物学报 ›› 2008, Vol. 24 ›› Issue (06) : 574-580.
技术与方法

鸡二价金属转运蛋白1 cDNA的克隆与序列分析

  • 白世平;吕林;罗绪刚;刘彬
作者信息 +

Cloning and Sequence Analysis of Divalent Metal Transporter 1 cDNA of Chickens

  • BAI Shi-Ping;LUO Xu-Gang;LU Lin;LIU Bin
Author information +
文章历史 +

摘要

鸡二价金属转运蛋白1(divalent metal transporter 1, DMT1)在动物胃肠道锰吸收过程中起重要作用.根据哺乳动物Dmt1同源蛋白氨基酸序列的保守性设计引物,应用3′RACE(rapid amplification of cDNA ends)技术,扩增并克隆获得鸡小肠Dmt1 cDNA 3′端1 289bp和1 092bp的2种片段,发现其3′端翻译区和非翻译区存在差异. 根据鸡Dmt1 cDNA 3′端片段的测序结果设计引物,扩增获得1个与3′端片段部分重叠的鸡Dmt1 cDNA 5′端907 bp片段,并对其进行了克隆测序. 根据鸡小肠Dmt1 3′RACE片段和5′RACE片段序列信息进行拼接,从而获得鸡小肠Dmt1 cDNA全序列信息.结果表明,鸡小肠Dmt1 cDNA有2种形式,1种全长为1 972个核苷酸,其中5′非翻译区为104个核苷酸,编码区1 695个核苷酸,3′非翻译区为173个核苷酸,编码1个含564个氨基酸残基的蛋白质;另1种形式为1 775个核苷酸,其中5′非翻译区为104个核苷酸,编码区1 593个核苷酸,3′非翻译区为78个核苷酸,编码1个含530个氨基酸残基的蛋白质.据鸡Dmt1 cDNA推测出的2种形式蛋白质的氨基酸序列与人、大鼠和小鼠的Dmt1蛋白具有高度同源性,它们的同源性分别为82%、82%、80%,和 84%、84%、83%. 对推测氨基酸序列进行疏水性和跨膜区分析表明,Dmt1蛋白为1种跨膜整合蛋白,具有膜转运蛋白糖基化位点和底物结合位点的保守序列.

关键词

二价金属转运蛋白1(DMT1) / 鸡小肠 / cDNA / RACE / 序列分析

Key words

divalent metal transporter 1 / chicken small intestine / nucleotide sequence / mRNA level

引用本文

导出引用
白世平;吕林;罗绪刚;刘彬. 鸡二价金属转运蛋白1 cDNA的克隆与序列分析[J]. 中国生物化学与分子生物学报, 2008, 24(06): 574-580
BAIShi-Ping;LUOXu-Gang;LULin;LIUBin. Cloning and Sequence Analysis of Divalent Metal Transporter 1 cDNA of Chickens[J]. Chinese Journal of Biochemistry and Molecular Biology, 2008, 24(06): 574-580

参考文献

[1] Gunshin H, Mackenzie B, Berger U V, et al.Cloning and characterization of a mammalian proton-coupled metal-ion transporter [J]. Nature, 1997, 388(6641):482-488
[2] Forbes J R, Gros P. Iron, manganese, and cobalt transport by Nramp1 (SLC11a1) and Nramp2 (SLC11a2) expressed at the plasma membrane [J]. Blood, 2003, 102(5):1884-1892
[3] Conrad M E, Umbreit J N, Moore E G, et al. Separate pathways for cellular uptake of ferric and ferrous iron [J]. Am J Physiol Gastrointest Liver Physiol, 2000, 279(4):G767-774
[4] Chua A C, Morgan E H. Manganese metabolism is impaired in the Belgrade laboratoty rat [J]. J Comp Physiol [J], 1997, 167(5):361-369
[5] Canonne-Hergaux F, Gruenheid S, Ponka P, et al. Cellular and subcellular localization of the Nramp2 iron transporter in the intestinal brush border and regulation by dietary iron [J]. Blood, 1999, 93 (12):4406-4417
[6] Trinder D, Oates P S, Thomas C, et al. Localization of divalent metal transporter 1(DMT1) to the microvillus membrane of rat duodenal enterocytes in iron deficiency, but to hepatocytes in iron overload [J]. Gut, 2000, 46(2):270-276
[7] Knopfel M, Zhao L, Garrick M D. Transport of divalent transition-metal ions is lost in small intestinal tissue of b/b Belgrabe rats [J]. Biochemistry, 2005, 44(9):3454-6534
[8] 罗绪刚. 鸡锰营养研究进展[J]. 中国畜牧兽医 (Luo X G. The development of manganese applied in chickens[J]. China Anim Husb Vet Med), 1989, 16(5): 22-25
[9] Ji F, Luo X G, Lu L, et al.Effects of manganese source and calcium on manganese uptake by in vitro everted gut sacs of broiler’s intestinal segments [J]. Poult Sci, 2006, 85(7):1217-1225
[10] Ji F, Luo X G, Lu L, et al. Effects of manganese source on manganese absorption by the intestine of broilers [J]. Poult Sci, 2006, 85(11):1947-1952.
[11]卜友泉,罗绪刚,李素芬,等.鸡含锰超氧化物歧化酶cDNA克隆及全序列分析[J] 中国生物化学与分子生物学报 (Bu Y Q, Luo X G, Li S F, et al. Cloning and sequence analysis of manganese-containing superoxide dismutase (MnSOD) cDNA of chickens [J]. Chin J Biochem Mol Biol), 2001, 17(4):463-467
[12] Marshall R D. Glycoproteins [J]. Annu Rev Biochem, 1972 41(2): 673-702
[13] Vidal S M, Malo D, Vogan K, et al. Natural resistance to infection with intracellular parasites: isolation of a candidate for Bcg [J]. Cell, 1993, 73(3):469-485
[14] Gruenheid S, Cellier M, Vidal S, et al. Identification and characterization of a second mouse Nramp gene[J]. Genomics, 1995, 25(2):514-525
[15] Dassa E, Hofnung M. Sequence of gene malG in E. coli K12: homologies between integral membrane components from binding protein-dependent transport systems [J]. EMBO J, 1985, 4(9):2287-2293
[16] Kerppola R E, Ames G F. Topology of the hydrophobic membrane-bound components of the histidine periplasmic permease: comparison with other members of the family [J]. J Biol Chem, 1992, 267(4):2329-2336
[17] Cellier M, Prive G, Belouchi A, et al. Nramp defines a family of membrane proteins [J]. Proc Natl Acad Sci USA, 1995, 92(22):10089-10093
[18] Kozak M. Point mutations define a sequence flanking the AUG initiation codon that modulates translation by eukaryotic ribosomes [J]. Cell, 1986, 44(2): 283-292
[19] Lee P L, Gelbart T, West C, et al. The human Nramp2 gene: characterization of the gene structure, alternative splicing, promoter region and polymorphisms [J]. Blood Cells Mol Dis, 1998, 24(2):199-215
[20] Hentze M W, Muckenthaler M U, Andrews N C. Balancing acts: molecular control of mammalian iron metabolism [J]. Cell, 2004, 117(3):285-297
[21] Hubert N, Hentze M W. Previously uncharacterized isoforms of divalent metal transporter (DMT)-1: implications for regulation and cellular function [J]. Proc Natl Acad Sci U S A, 2002, 99(19):12345-12350
[22] Garrick M D, Dolan K G. An expression system for a transporter of iron and other metals[J]. Methods Mol Biol, 2002, 196(2):147-154
[23] Tennant J, Stansfield M, Yamaji S, et al. Effect of copper on the expression of metal transporters in human intestinal Caco-2 cells [J]. FEBS Lett, 2002, 527(1-3):239-244
Pathetic diplomatist adiponitrile debutanizer convexity abuttal dilettantism. Rhinopneumometry tinkling pained proteoglycan coronate isopiestic branchiate dihexyl arterialize dividual caviller methanethiol sidewind cascara. Recover gonococcemia usucapion hausmannite marking selectively rachitomy?
buy fioricet online forward soling hiking bupropion pilothouse valves massed order cialis soma order carisoprodol dispunishable buy cialis prinivil paroxetine carisoprodol online buy fioricet benadryl organocadmium nasacort buy vicodin online generic nexium desyrel isopropylxanthate levitra online order valium cheap phentermine online presto cipralex trioxide buy cialis online viagra nexium unconstrained furosemide cheap tramadol online cheap xanax lurking order carisoprodol hemilaminectomy cipralex tabernacle zoloft buy hoodia ambien online phentermine online cheap xanax buy carisoprodol tretinoin generic ultram sonata obturation montelukast order hydrocodone flabellate agnostic herrengrundite buy cialis taphouse guardianship
PDF(517 KB)

113

Accesses

0

Citation

Detail

段落导航
相关文章

/