Paddy rice zinc finger protein gene and its coded protein
A zinc finger protein and gene coding technology, applied in the field of genetic engineering, can solve the problems that the relationship between WZF1 stress response has not been found, and the function description of rice C2H2 zinc finger protein gene has not yet been described.
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Embodiment 1
[0034] Molecular Cloning of Rice Zinc Finger Protein Gene OsZFP18
[0035] The rice variety "Jiucaiqing" (japonica rice local variety in the Taihu Lake Basin, a strong salt-tolerant variety) was selected. After the rice seedlings grew to the 3-leaf stage, they were treated with 140mM NaCl, and the leaves were immediately taken out and frozen in liquid nitrogen after 12 hours. . Take part of the leaves, grind them with a mortar, add them into a 1.5mLEP tube filled with lysate, oscillate fully, and then transfer them into a glass homogenizer. After homogenization, transfer to 1.5mL EP tube, and extract total RNA (TRIzolReagents, Invitrogen, USA). The quality of total RNA was identified by formaldehyde denaturing gel electrophoresis, and then the RNA content was determined on a spectrophotometer. The GenBank rice genome database was searched with the EST sequence S121 from the rice salt-induced cDNA library, and the full-length 883bp sequence of the rice C2H2 zinc finger protei...
Embodiment 2
[0039] Sequence information and characteristic analysis of OsZFP18
[0040]The full length of OsZFP18 of the present invention is 883bp, and its open reading frame is located at positions 53-568. DNAssist software analysis showed that OsZFP18 encoded a total of 171 amino acids, which contained 2 typical Cys2 / His2 zinc finger structures, and the Cys-Xaa-Cys and His-Xaaa-His regions were composed of 12 mainly acidic or hydrophobic amino acids The coordination structure plus the hydrophobic core formed by Phe and Leu determine the basic structure and shape of the zinc finger. Both zinc finger structures contain the conserved structure of QALGGH. By Blast program comparison, it was found that the amino acid similarities between the encoded protein of OsZFP18 and soybean zinc finger protein SCOF-1, wheat WZF1, and Arabidopsis STZ were 33%, 36%, and 30%, respectively. Its pI=5.19 and MW=17.9 KDa were estimated using Compute pI / MW software. Searching the existing rice genome (incl...
Embodiment 3
[0042] Expression study of OsZFP18
[0043] Using the primers at both ends of OsZFP18 designed in Example 1 to carry out semi-quantitative RT-PCR analysis of the expression of rice seedling shoots and underground parts, using the expression of rice actin gene Rac1 (McElroy et al, 1990) as an internal reference, the results showed that rice seedlings OsZFP18 The expression of was significantly enhanced under 140mM NaCl treatment for 6h, and reached the highest level at 24h. The grayscale scanning analysis showed that the expression level was about 10 times that of the control.
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