Cloning, Express and the Construction of Yeast Shuttling Expression Plasmid of Silkworm BmNaPi Gene

doi:10.7668/hbnxb.2011.03.011

Abstract

Abstract: Na(+)-dependent phosphate cotransporter(NaPi) mediate the transport and metabolism of inorganic phosphate.Phosphate is essential for silkworm and the homologous gene of silkworm,BmNaPi,is obtained through bioinfomatics analysis and molecular biological experiment.The gene is located on 8th chromosome,1 437 bp long,and encodes 478 amino acids,with a signal peptide in the N-termi and 8 transmembrane domains inside.A comparison of BmNaPi with homologous protein AAF57968,XP_001602874,EAA00281,XP_393759,XP_001950194 from other sources revealed that their amino acids share above 70% identity.It is shown by RT-PCR experiment that the BmNaPi has expression in blood,integument,gonad and head,while no expression in midgut,silk gland,malpighian tubule and fat body among 9 tissues detected on day 3 of the fifth instar larvae.Then yeast shuttling expression plasmid pGBKT7-BmNaPi was successfully constructed.

Key words: Bombyx mori, BmNaPi, Clone, Tissue expression, Yeast shuttling expression plasmid

CLC Number:

S881.2

KONG Wei-qing, YANG Jin-hong. Cloning, Express and the Construction of Yeast Shuttling Expression Plasmid of Silkworm BmNaPi Gene[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(3): 51-55. doi: 10.7668/hbnxb.2011.03.011.

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References

[1] FeildJA,ZhangL,BrunKA,et al.Cloning and fnctional characterization of a sodium-dependent phosphate transporter expressed in human lung and small intestine[J].BBRC,1999,258(3):578-582.

[2] Kiyamova,Gryshkova V,Ovcharenko G,et al.Development of monoclonal antibodies specific for the human sodium-dependent phosphate Co-transporter NaPi2b[J].Hybridoma,2008,27(4):277-284.

[3] 张越,杨洋,安继红,等.磷酸钠协同转运蛋白基因SLC34A2的真核表达重组载体的构建及鉴定[J].中国老年学杂志,2008,28(6):564-566.

[4] Segawa H,Aranami F,Kaneko I,et al.The roles of Na/Pi-Ⅱ transporters in phosphate metabolism[J].Bone,2009,45:2-7.

[5] Hu Q,Izumi S,Miyazawa H,et al.Mutations in the SLC34A2 gene are associated with pulmonary alveolar microlithiasis[J].Am J Respir Crit Care Med,2007,175(3):263-268.

[6] Bai L Q,Collins J F,Ghishan F K.Cloning and characterization of a type Ⅲ Na-dependent phosphate cotransporter from mouse intestine[J].Am J Physiol Cell Physiol,2000,279:C1135-C1143.

[7] Lorenz-Depiereux B,Benet-Pages A,Eckstein G.Hereditary hypophosphatemic rickets with hypercalciuria is caused by mutations in the sodium-phosphate cotransporter gene SLC34A3[J].Am J Hum Genet,2006,78(2):193-201.

[8] Giral H,Caldas Y,Sutherland E,et al.Regulation of rat intestinal Na-dependent phosphate transporters by dietary phosphate[J].Am J Physiol Renal Physiol,2009,297:F1466-F1475.

[9] Villa-Bellosta R,Ravera S,Sorribas V,et al.The Na +-Pi cotransporter PiT-2 (SLC20A2) is expressed in the apical membrane of rat renal proximal tubules and regulated by dietary Pi[J].Am J Physiol Renal Physiol,2009,296:F691-F699.

[10] Li X W,Yang H Y,Giachelli C M.Role of the sodiumdependent phosphate cotransporter,Pit-1,in vascular smooth muscle cell calcification[J].Circulation Research,2006,98:905-912.

[11] XIA Q Y,ZHOU Z Y,LU C,et al.A draft sequence for the genome of the domesticated silkworm (Bombyx mori)[J].Science,2004,306:1937-1940.

[12] Duan J,Li R Q,Cheng D J,et al.SilkDB v2.0:aplatform for silkworm (Bombyx mori) genome biology[J].Nucl Acids Res,2009:1-4.

[13] MITA K,MORIMYO M,OKANO K,et al.The construction of an EST database for Bombyx mori and its application[J].PNAS,2003,100(24):14121-14126.

[14] 程道军,夏庆友,周泽扬,等.家蚕cDNA文库构建及大规模EST测序[J].蚕业科学,2003,29(4):335-339.

[15] Virkki L V,Biber J,Murer H,et al.Phosphate transporters:a tale of two solute carrier families[J].Am J Physiol Renal Physiol,2007,293:F643-F654.

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