Cloning of a NaCl-induced fructose-1, 6-diphosphate aldolase cDNA from Dunaliella salina and its expression in tobacco.
Using Rapid Amplification of cDNA ends (RACE) technique, the full-length cDNA encoding a NaCl-induced fructose-1, 6-diphosphate aldolase (DsALDP) was obtained. It was shown that the DsALDP had a relatively high homology (66%-73%) to chloroplast fructose-1, 6-diphosphate aldolase (AldP) in many plants according to their amino acid sequences. The phylogenetic analysis further confirmed that AldP in alga is the nearest to DsALDP. As to its expression pattern, DsALDP was de novo synthesized by NaCl induction. Its expression level was significantly changed with inducing time. After the selected DsALDP cDNA subcloned into a binary vector pBI121, the new construct was introduced into tobacco by Agrobacterium tumefaciens. The results of Southern blot and RT-PCR analysis of four transgenic T1 plants indicated that DsALDP was integrated into genome of these transgenic plants and effectively expressed. Aldolase activities have been detected in T1-1, T1-2 and T1-3 plants by bioassay under 100-200 mmol/L NaCl. It was also observed that proline contents in them were differentially increased.
Zhang
Xiaoning
X
Institute of Genetics, State Key Laboratory of Genetic Engineering, Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China.
Lin
Changfa
C
Chen
Huoying
H
Wang
Hao
H
Qu
Zhicai
Z
Zhang
Hongwei
H
Yao
Jianhong
J
Shen
Daleng
D
D020418:Algal Proteins / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D000460:Chlorophyta / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D048028:DNA, Algal / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics*; D015870:Gene Expression; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000235:genetics; D055051:Salt-Tolerant Plants / Q000201:enzymology / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D012965:Sodium Chloride / Q000494:pharmacology; D014026:Nicotiana / Q000201:enzymology* / Q000235:genetics*; D014882:Water-Electrolyte Balance / Q000235:genetics / Q000502:physiology
D020418:Algal Proteins; D048028:DNA, Algal; D018076:DNA, Complementary; D011994:Recombinant Proteins; D012965:Sodium Chloride; D005634:Fructose-Bisphosphate Aldolase