Plant nonspecific lipid transfer proteins (nsLTPs) are widely distributed through plant kingdom and are characterized by the presence of a central hydrophobic cavity, suitable for binding various hydrophobic molecules...Plant nonspecific lipid transfer proteins (nsLTPs) are widely distributed through plant kingdom and are characterized by the presence of a central hydrophobic cavity, suitable for binding various hydrophobic molecules. Despite extensive research on nsLTP in different plant species, mostly angiosperm, and the great diversity of physiological processes in which they seem to be involved, their exact functions still remain unclear. Also, very limited experimental data are available on nsLTP in gymnosperm. In this study, we report for the first time on the molecular cloning of nsLTP, from Pinus sylvestris L.(PsLTP1, GenBank accession JN980402.1) and the expression pattern of PsLTP1 during ontogenesis and in response to environmental stress conditions. Total RNA from roots of 7-day old pine seedlings was used to isolate the cDNA clone, corresponding to Scots pine lipid transfer protein. The open reading frame of PsLTP1 consists of 372 bp encoding a protein of 123 amino acids. Amino acid sequence alignment revealed that mature PsLTP1 shares high level of similarity with nsLTP from other conifers and with well-studied nsLTPs from angiosperms. The PsLTP1 contains a 27-amino-acid N-terminal signal sequence and presents all the features of a plant nsLTP. Amino acid comparison analysis and 3D structure prediction showed that PsLTP1 is a type 1 nsLTP. The results of the expression analysis of Scots pine PsLTP1 gene revealed that its transcripts accumulate in actively growing tissues. Furthermore, transcription of PsLTP1 was upregulated in response to cold and salt treatments, and downregulated during acidic, osmotic and water stresses.展开更多
通过PCR方法从水稻(O ry za sa tiva)染色体基因组中扩增一个编码非特异性脂质转移蛋白(nonspec ific lip id transfer prote in,nsLTP)类似蛋白的基因(OSd ir),将该基因片段克隆到硫氧还蛋白融合表达载体pET 32a(+)中后,在BL 21(DE 3)t...通过PCR方法从水稻(O ry za sa tiva)染色体基因组中扩增一个编码非特异性脂质转移蛋白(nonspec ific lip id transfer prote in,nsLTP)类似蛋白的基因(OSd ir),将该基因片段克隆到硫氧还蛋白融合表达载体pET 32a(+)中后,在BL 21(DE 3)trxB-宿主菌中实现了融合蛋白的高表达,表达产物为包涵体。利用尿素溶解包涵体并复性后,再通过Sepharose N i2+-che lating fast flow柱亲和纯化融合蛋白,得到重组O SD IR蛋白。展开更多
基金supported by grants from the Ministry of Education and Science of Ukraine(0116U003593)grant from cieA3(Campus de Excelencia Internacional Agroalimentario)-UCO,Spain
文摘Plant nonspecific lipid transfer proteins (nsLTPs) are widely distributed through plant kingdom and are characterized by the presence of a central hydrophobic cavity, suitable for binding various hydrophobic molecules. Despite extensive research on nsLTP in different plant species, mostly angiosperm, and the great diversity of physiological processes in which they seem to be involved, their exact functions still remain unclear. Also, very limited experimental data are available on nsLTP in gymnosperm. In this study, we report for the first time on the molecular cloning of nsLTP, from Pinus sylvestris L.(PsLTP1, GenBank accession JN980402.1) and the expression pattern of PsLTP1 during ontogenesis and in response to environmental stress conditions. Total RNA from roots of 7-day old pine seedlings was used to isolate the cDNA clone, corresponding to Scots pine lipid transfer protein. The open reading frame of PsLTP1 consists of 372 bp encoding a protein of 123 amino acids. Amino acid sequence alignment revealed that mature PsLTP1 shares high level of similarity with nsLTP from other conifers and with well-studied nsLTPs from angiosperms. The PsLTP1 contains a 27-amino-acid N-terminal signal sequence and presents all the features of a plant nsLTP. Amino acid comparison analysis and 3D structure prediction showed that PsLTP1 is a type 1 nsLTP. The results of the expression analysis of Scots pine PsLTP1 gene revealed that its transcripts accumulate in actively growing tissues. Furthermore, transcription of PsLTP1 was upregulated in response to cold and salt treatments, and downregulated during acidic, osmotic and water stresses.
文摘通过PCR方法从水稻(O ry za sa tiva)染色体基因组中扩增一个编码非特异性脂质转移蛋白(nonspec ific lip id transfer prote in,nsLTP)类似蛋白的基因(OSd ir),将该基因片段克隆到硫氧还蛋白融合表达载体pET 32a(+)中后,在BL 21(DE 3)trxB-宿主菌中实现了融合蛋白的高表达,表达产物为包涵体。利用尿素溶解包涵体并复性后,再通过Sepharose N i2+-che lating fast flow柱亲和纯化融合蛋白,得到重组O SD IR蛋白。
基金supported by grants from the National Science and Technology Pillar Program of China(2007BAD78B03)the “Eleventh-Five” Key Project of Sichuan ProvinceChina(07SG111-003-1)
