Application of cds sequence of rice transcription factor os11g35030.1 gene
A rice transcription factor and sequence technology, applied in the field of genetic engineering, can solve problems such as the weakening of hybrid breeding advantages, and achieve the effect of improving rice grain traits
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0032] Example 1 Obtaining CDS sequence of Os11g35030.1 gene and construction of plant expression vector
[0033] 1 Acquisition of CDS sequence of Os11g35030.1 gene
[0034] Find the Os11g35030.1 gene in the plant transcription factor database (http: / / rice.plantbiology.msu.edu / analyses_search_locus.shtml), design PCR amplification primers according to its CDS sequence, forward primer F 5'-CAAAAAAGCAGGCTTCATGCTGAGCTCTTGTGG-3' and reverse primer R5'-CAAGAAAGCTGGGTCTTAGAGAAGTGTTGGGAC-3'). The complete CDS sequence of the Os11g35030.1 gene (Seq ID No.1) was obtained by PCR using the total cDNA of wild-type Nipponbare 'kitaake' rice as a template and using primers F and R.
[0035] 2 Construction of plant expression vectors
[0036] The CDS sequence of the rice transcription factor Os11g35030.1 gene was constructed downstream of the four transcription factor activation motif VP16 coding genes (SEQ ID No.4) through the Gateway system.
[0037] 2.1 Cloning the above PCR product in...
Embodiment 2
[0047] The acquisition of embodiment 2 transgenic rice plants
[0048] Take the mature seeds of rice 'kitaake', shell them manually or mechanically, select the plump, smooth and spot-free seeds and inoculate them on the induction medium for induction culture after being sterilized. The rice callus with good appearance and good growth was selected as the recipient material, and ubi:VP64-Os11g35030.1 was transferred into the rice callus by the Agrobacterium-mediated method, with 100 μM acetosyringone and OD value 0.7 AAM culture solution of Agrobacterium for transformation, put the callus soaked in the transformation solution on the co-culture medium for co-cultivation, culture in the dark at 25°C for 3 days, then place it on the screening medium for about 30 days, subculture every 10 days once. Then, the screened out resistant calli were transferred to the differentiation medium for differentiation for about 20 days, and subcultured every 10 days. The resistant callus that di...
Embodiment 3
[0054] Identification of embodiment 3 transgenic positive strains
[0055] In order to detect the overexpression of ubi:VP64-Os11g35030.1 gene in T2 transgenic rice, primers were designed according to the carrier ubi:VP64-Os11g35030.1 (forward primer F5'-GTGGGGACAAGTTTGTACAAAAAAGCAGGCTTC-3' and reverse primer R 5' -GTGGGGACCACTTTGTACAAGAAAGCTGGGTC-3') was detected by PCR, and the amplified product was subjected to agarose gel electrophoresis, and obvious specific bands appeared. Among them, WT means wild-type rice 'kitaake', V0257H-07 and V0257H-14 mean VP64-Os11g35030.1 transgenic rice line ( image 3 ).
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 


