A kind of filial piety bamboo transcription factor bmbzip62 gene and its application
A transcription factor and gene technology, which is applied to the filial bamboo transcription factor BmbZIP62 gene and its application field, can solve the problem that the new function needs to be verified and so on.
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Embodiment 1
[0025] Example 1: Acquisition of gene encoding BmbZIP62 transcription factor in filial bamboo
[0026] 1. RNA extraction
[0027] The extraction material was bamboo shoots of filial piety bamboo. 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 Primer 1μL, 10μL 2×TS Reaction Mix, 1μL TransScript RT / RI Enzyme Mix, 1μL 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.
[0030] 3. PCR am...
Embodiment 2
[0033] Example 2: Functional Analysis of BmbZIP62 Transcription Factor of Filial Bamboo
[0034] 1. Sequence comparison analysis
[0035] In order to analyze the genetic relationship between BmbZIP62 of filial bamboo and bZIP transcription factors in model plants Arabidopsis and rice, a phylogenetic tree of BmbZIP62 and bZIP transcription factors in other species was established using MEGA7. result( figure 2 ) indicated that BmbZIP62 had the closest relationship with rice OsbZIP62 and OsbZIP29 transcription factors.
[0036] The amino acid sequence homology of BmbZIP62 and its closest relatives OsbZIP62 and OsbZIP29 was analyzed by DNAMAN8. Analysis results such as image 3 As shown, the sequence similarity between BmbZIP62 and OsbZIP62 is 61.27%, and the sequence similarity between BmbZIP62 and OsbZIP29 is 47.97%.
[0037] 2. Arabidopsis transgene analysis
[0038] 2.1 Vector construction
[0039] The PCR product whose sequencing sequence was consistent with the transc...
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