|Table of Contents|

Cloning and Expression Analyses of Sporamin Family Genes in Sweet Potato(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

2013 02
Research Field:
Publishing date:


Cloning and Expression Analyses of Sporamin Family Genes in Sweet Potato
LAI Xianjun GU Yinghong WANG Haiyan ZHANG Yizheng
(Sichuan Key Laboratory of Molecular Biology & Biotechnology, College of Life Sciences, Sichuan University, Chengdu 610064, China)
sweet potato sporamin gene cloning sequence analysis digital gene expression profiling protease inhibitor
Q946.1 : Q78

In order to study the structure and function of sporamin family genes in sweet potato, the transcriptome database of this plant constructed in our lab were analyzed to screen sporamin contigs and appropriate primers designed to amplify sporamin genes, which were then cloned and sequenced. All together 61 sporamin contigs were found in the transcriptome database, including 31 contigs in sporamin A sub-family and 30 contigs in sporamin B sub-family. Six sporamin A genes and 10 sporamin B genes were cloned and sequenced. ORF of all sporamin A genes were 660 bp in length with identity more than 95% among all sporamin A genes. ORF of sporamin B genes had a total length of 651 bp with identity more than 98% among sporamin B genes. Homology between the two sub-families was 84%. The results of digital gene expression profile analysis showed that both sporamin A and B had the highest level of transcription in the initial tuberous roots and harvest tuberous roots, comparatively low transcription in the expanding tuberous roots, and very little in leaves and stems. Analysis of conserved sequence of sporamin genes and homology comparison displayed that the proteins contained a soybean trypsin inhibitor conserved sequence with more than 170 amino acids belonging to the Kunitz-type protease inhibitor superfamily, indicating that the sporamin has the function of protease inhibitors. Fig 9, Tab 3, Ref 33


1 Maeshima M, Sasaki T, Asahi T. Characterization of majorproteins in sweet potato tuberous roots [J]. Phytochemistry, 1985, 24: 1899-1902
2 Hattori T, Nakamura K, Yoshida. Structural relationship among the members of a multi-gene family coding for the sweet potato tuberous root storage protein [J]. Plant Mol Biol, 1989, 13: 563-572
3 Murakami S, Hattori T, Nakamura K. Structural differences in full-length cDNAs for two classes of sporamin, the major soluble protein of sweet potato tuberous roots [J]. Plant Mol Biol, 1986, 7: 343-355
4 Wettstein D, Chua NH. Plant Molecular Biology [M]. New York: Plenum Press, 1987
5 Conlan RS, Griffiths LA, Napier JA, Shewry PR, Mantell S, Ainsworth C. Isolation and characterisation of cDNA clones representing the genes encoding the major tuber storage protein (dioscorin) of yam (Dioscorea cayenensis Lam.) [J]. Plant Mol Biol, 1995, 28 (3): 369-380
6 Yang J, Barr LA, Fahnestock SR, Liu ZB. High yield recombinant silk-like protein production in transgenic plants through protein targeting [J]. Transgenic Res, 2005, 14: 313-324
7 Woofe JA. Sweet Potato, An Untapped Food Resource [M]. New York: Cambridge University Press, 1992
8 Yeh KW, Chen JC, Lin MI, Chen YM, Lin CY. Functional activity of sporamin from sweet potato (Ipomoea batatas Lam.): a tuber storage protein with trypsin inhibitory activity [J]. Plant Mol Biol, 1997, 33: 565-570
9 Yeh KW, L in MI, Tuan SJ, Chen YM, Lin CJ, Kao SS. Sweet potato (Ipomoea batatas) trypsin inhibitors expressed in transgenic tobacco plants confer resistance against Spodoptera litura [J]. Plant Cell Rep, 1997, 16 (10): 696-699
10 Koiwa H, Shade RE, Zhu-Salzman K, D’Urzo MP, Murdock LL, Bressan RA, Hasegawa PM. A plant defensive cystatin (soyacystatin) targets cathepsin L-Like digestive cysteine proteinases (DvCALs) in the larval midgut of western corn rootworm (Diabrotica virgifera) [J]. FEBS Lett, 2000, 471: 67-70
11 Hong YF, Liu CY, Cheng KJ, Hour AL, Chan MT, Tseng TH, Chen KY, Shaw JF, Yu SM. The sweet potato sporamin promoter confers high-level phytase expression and improves organic phosphorus acquisition and tuber yield of transgenic potato [J]. Plant Mol Biol, 2008, 67: 347-361
12 邱琳, 董衡, 黄鹂, 曹家树. 甘薯sporamin蛋白的功能及其在抗虫转基因植物中的应用[J]. 中国细胞生物学学报, 2011, 33 (6): 692-698 [Qiu L, Dong H, Huang L, Cao JS. The function of sweet potato’s special tuber storage protein and its application in engineering plant research [J]. Chin J Cell Biol, 2011, 33 (6): 692-698]
13 Hurley BA, Tran HT, Marty NJ, Park J, Snedden WA, Mullen RT, Plaxton WC. The dual-targeted purple acid phosphatase isozyme At PAP26 is essential for efficient acclimation of Arabidopsis to nutritional phosphate deprivation [J]. Plant Physiol, 2010, 7 (153): 1112-22
14 陈克贵, 王海燕, 张义正. 甘薯贮藏蛋白(sporamin) A基因编码区克隆及序列同源性分析[J]. 中国生物化学与分子生物学报, 2000, 16 (1): 41-45 [Chen KG, Wang HY, Zhang YZ. Cloning and homologous analysis of sporamin A gene coding region sequence of Ipomoea batatas [J]. Chin J Biochem Mol Biol, 2000, 16 (1): 41-45]
15 封树梅, 张义正. 甘薯储藏蛋白(sporamin) A基因在大肠杆菌中的表达研究[J]. 四川大学学报(自然科学版), 2000, 37 (3): 446-450 [Feng SM, Zhang YZ. Expression of sporamin A gene of sweet potato in Escherichia coli [J]. Jf Sichuan Univ (Nat Sci Ed), 2000, 37 (3): 446-450]
16 Tao X, Gu YH, Wang HY, Zheng W, Li X, Zhao CW, Zhang YZ. Digital gene expression analysis based on integrated de novo transcriptome assembly of sweet potato [Ipomoea batatas (L.) Lam.] [J]. PLoS ONE, 2012, 7 (4): e36234
17 Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual [M]. 2nd ed. New York: Cold Spring Harbor Laboratory Press, 1989
18 Loebenstein G. Origin, distribution and economic importance [M]. The Sweetpotato. Springer Netherlands, 2009: 9-12
19 Trapnell C, Pachter L, Salzberg SL. Discovering splice junctions with RNA-Seq [J]. Bioinformatics, 2009, 25 (9): 1105-1111
20 程龙军, 郭得平, 葛红娟. 甘薯块根特异蛋白——sporamin的研究进展[J]. 植物学通报, 2001, 18 (6): 672-677 [Cheng LJ, Guo DP, Ge HJ. The special proteins in sweet potato tuber-sporamin [J]. Chin Bull Bot, 2001, 18 (6): 672-677]
21 Hattori T, Nakamura K. Genes coding for the major tuberous root protein of sweet potato: identification of putative regulartory sequence in the 5’ upstream region [J]. Plant Mol Biol, 1988, 22: 417-426
22 Chen JC, Chen YM, Yeh KW. Isolation and characterization of new sporamin gene members from sweet potato (Imomoea batatas Lam.) [J]. Taiwania, 1997, 42: 34-42
23 Ohta S, Hattori T, Morikami A. High-level expression of a sweet potato sporamin gene promoter β-glucuroidase (GUS) fusion gene in the stems of transgenic tobacco plants is conferred by multiple cell type-specific regulatory elements [J]. Mol Gen Genet, 1991, 225: 369-378
24 Marchler-Bauer A, Lu S, Anderson JB, Chitsaz F, Derbyshire MK, Weese-Scott CD, Fong JH, Geer LY, Geer RC, Gonzales NR, Gwadz M, Hurwitz DI, Jackson JD, Ke Z, Lanczycki CJ, Lu F, Marchler GH, Mullokandov M, Omelchenko MV, Robertson CL, Song JS, Thanki N, Yamashita RA, Zhang D, Zhang N, Zheng C, Bryant SH. CDD: a Conserved Domain Database for the functional annotation of proteins [J]. Nucleic Acids Res, 2011, 39D: 225-229
25 Hattori T, Nakagawa T, Maeshima M. Molecular cloning and nucleotide sequence of cDNA for sporamin, the major soluble protein of sweet potato tuberous roots [J]. Plant Mol Biol, 1985, 5: 313-320
26 Nakamura K, Matsushita K, Mukumoto F. Precursor to sweet potato sporamin in transformed tobacco cell line BY-2 [J]. J Exp Bot, 1993, 44: 331-338
27 Matsuoka K, Matsuomoto S, Hattori T. Vacuolar targeting and post-translational processing of the precursor to the sweet potato tuberous root storage protein in heterologous plant cells [J]. J Biol Chem, 1990, 265: 19750-19757
28 Arnold K, Bordoli L, Kopp J, Schwede T. A web-based environment for protein structure homology modelling [J]. Bioinformatics, 2006, 22: 195-201
29 Schwede T, Kopp J, Guex N, Peitsch MC. An automated protein homology-modeling server [J]. Nucleic Acids Res, 2003, 31: 3381-3385
30 Guex N, Peitsch MC. An environment for comparative protein modelling [J]. Electrophoresis, 1997, 18: 2714-2723
31 Yao PL, Hwang MJ, Chen YM, Yeh KW. Site-directed mutagenesis evidence for a negatively charged trypsin inhibitory loop in sweet potato sporamin [J]. FEBS Lett, 2001, 496: 134-138
32 Huang GJ, Sheu MJ, Chen HJ, Chang YS, Lin YH. Growth inhibition and induction of apoptosis in NB4 promyelocytic leukemia cells by trypsin inhibitor from sweet potato storage roots [J]. J Agric Food Chem, 2007, 55: 2548-2553
33 Yao J, Qian CJ. Sporamin induce apoptosis in human tongue carcinoma cells by down-regulating Akt/GSK-3 signaling [J]. Fundam Clin Pharmacol, 2011, 25 (2): 229-236


Last Update: 2013-05-02