Ibtip1 gene from sweet potato root and promoter thereof

A sweet potato and gene technology, applied in genetic engineering, plant gene improvement, peptide source, etc., can solve the unreported sweet potato TIP gene and other problems

Inactive Publication Date: 2009-05-06
KOREA RES INST OF BIOSCI & BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the TIP gene in sweet potato has never been reported

Method used

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  • Ibtip1 gene from sweet potato root and promoter thereof
  • Ibtip1 gene from sweet potato root and promoter thereof
  • Ibtip1 gene from sweet potato root and promoter thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Replication of sweet potato IbTIP1 gene

[0075] The fibrous root tissue of sweet potato (Ipomoea batatas (L.) Lam.cv.White star) was dried for 6 hours and then isolated by CTAB method (Kim and Hamada, Biotechnol Lett 27, 1841-1845, 2005) RNA. mRNA was isolated using the Poly(A) Tract mRNA Isolation System (Promega), and a sweet potato dried-treated root cDNA library was prepared using the SMART cDNA library (library) construction kit (Clontech). The first titer of the library shows about 1.5×10 6 pfu / ml value. 983 kinds of EST clones were secured by analysis based on base sequences extracted at random.

[0076] The nucleotide sequence of the 5' portion of the cDNA was determined using T3 primers, and the information and data of related genes were confirmed by analyzing the Blast database of NCBI (www.ncbi.nlm.gov). Among them, by analyzing the data, the candidate gene that may show specific expression in dryness and roots was copied, that is, the TIP (tonoplast in...

Embodiment 2

[0078] The expression analysis of the IbTIP1 gene of each tissue of sweet potato

[0079]In order to analyze the expression state of each tissue of the sweet potato moisture transport channel protein gene IbTIP1 of the present invention isolated from the dried sweet potato root, RT-PCR was performed. Whole RNA was extracted from sweet potato tissues (leaves, stems, fibrous roots, thick roots, and storage roots) using the CTAB method (Kim and Hamada 2005), and then 2.5 μg of total RNA was used to synthesize cDNA using the ImPro-H Reverse Transcrirption System cDNA synthesis kit (Promega Corporation) Synthesize cDNA. The expression state of the IbTIP1 gene was investigated using IbTIP1 gene-specific primers and using Ex-Taq DNA polymerase (TaKaRa). Base sequences (Forward: 5'-GTCACGTAAACCCTGCTGTCAC-3': SEQ ID NO: 4) and (Reverse: 5'-GCTAGAATGTTGGCACCCACTA-3': SEQ ID NO: 5) were used as IbTIP1-specific primers.

[0080] As a result, the IbTIP1 gene was strongly expressed in fi...

Embodiment 3

[0082] Expression Analysis of the IbTIP1 Gene Based on Drying Treatment and Hormone Treatment

[0083] In order to analyze the degree of response of the IbTIP1 gene of the present invention according to the drying treatment, the leaves and fibrous root tissues of sweet potato were dried for different times (0, 1, 2, 4, 8, 16, 24 hours) and then used in the Examples The method in 2 was used to isolate RNA, and then synthesize cDNA separately. RT-PCR was performed using the AccelTaq Premix Kit (Gedocs) using the IbTIP1 gene-specific primers used in Example 2 above.

[0084] As a result, the IbTIP1 gene in the leaf tissue began to increase gradually after 1 hour after the drying process began, and showed an expression peak in 4-8 hours, and then showed a reduced state ( Figure 4a ). In contrast, no increase or decrease in the expression of IbTIP1 based on the drying treatment was observed in the fibrous root tissue. This may be due to the high expression level of IbTIP1 gene...

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Abstract

The present invention relates to an IbTIP1 gene from Ipomoea batatas L.Lam root and a promoter thereof. More particularly, the invention relates to IbTIP1 gene which is super expressed at Ipomoea batatas L.Lam root and has expression induced by various stresses and comes from Ipomoea batatas L.Lam root, and the promoter thereof. The IbTIP1 gene and the promoter according to the invention is suitable for the development of environment stress resistant plant suitable for areas with poor condition, transformation Ipomoea batatas L.Lam which can produce useful raw materials of starch, functional protein, etc., or the development of environment-friendly energy-replacing crop used for producing bioethanol.

Description

technical field [0001] The present invention relates to the IbTIP1 gene derived from sweet potato roots and its promoter (promoter), more specifically, to the IbTIP1 gene derived from IbTIP1 gene and its promoter in sweet potato root. Background technique [0002] Sweet potato (Ipomoea batatas L.Lam) can not only be cultivated in relatively poor soil, but also has a yield of about 22 tons per hectare. It is a representative root crop used as food and livestock feed. In particular, about 70% by weight of dried sweet potato is composed of starch, so it has been used as a raw material crop for ethanol, and has recently been used as an environment-friendly alternative energy crop for bioethanol production. [0003] Recently, global environmental problems and food problems have been raised due to rapid industrialization and population increase. Therefore, the development of environmentally disaster-resistant crops that are suitable for survival in desertified areas, polluted are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N15/82C12N15/11
CPCC07K14/415C12N15/63C12N15/8227C12N15/8257
Inventor 郭尚洙李幸顺金且英金善衡梁暻实宋完根
Owner KOREA RES INST OF BIOSCI & BIOTECH
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