Bambusa multiplex transcription factor BmMYB83 gene and application thereof
A transcription factor and gene technology, applied in the filial piety bamboo transcription factor BmMYB83 gene and its application field, can solve problems such as almost blank research
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Embodiment 1
[0025] Example 1: Acquisition of gene encoding transcription factor of filial piety bamboo BmMYB83
[0026] 1. RNA extraction
[0027] The extraction material is bamboo shoots of filial piety bamboo. The RNA was extracted using the Yuanping Hao RNAPure Ultra-Pure Rapid Extraction Kit, extracted according to the operating instructions, and stored at -80°C for later use
[0028] 2. cDNA first-strand synthesis and reverse transcription PCR
[0029] The total RNA was reverse-transcribed into cDNA using the Whole Type Gold (Beijing) one-step cDNA synthesis kit, following the operating instructions. Reaction system and reaction conditions are: 2 μg prepared total RNA, Anchored Oligo(dT) 18 Primer1μL, 10μL of 2×TS Reaction Mix, 1μL of TransScript RT / RI Enzyme Mix, 1μL of gDNARemover, add RNase-free Water to make up to 20μL, mix gently and centrifuge. Reverse transcription at 42°C for 30 minutes in a PCR instrument, 5s at 85°C; cool on ice; store at -20°C after centrifugation.
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Embodiment 2
[0033] Example 2: Functional Analysis of BmMYB83 Transcription Factor in Filial Bamboo Bamboo
[0034] 1. Sequence comparison analysis
[0035] In order to analyze the genetic relationship between BmMYB83 and MYB transcription factors of other species, a phylogenetic tree of BmMYB83 and MYB transcription factors of other species was established using MEGA7. result( figure 2 ) indicated that BmMYB83 had the closest relationship with OsMYB83 and OsMYB46 in the model plant rice, and belonged to the same cluster.
[0036] The homology of BmMYB83 with closely related MYB transcription factors in other species was analyzed by DNAMAN. result( image 3 ) showed that BmMYB83 had a high homology with OsMYB83 in rice, and the sequence similarity was 58.84%. The sequence similarities with rice OsMYB46 and Arabidopsis AtMYB83 were 39.44% and 38.39%, respectively.
[0037] 2. Rice transgenic analysis
[0038] 2.1 Vector construction
[0039] The correctly sequenced PCR product was l...
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