Phenotypes

Vacuolar H+/Ca2+ transporter; play a central role in Ca2+ and metal sequestration into the vacuole, localized in the tonoplast; predominately expressed in leaves. Altered plant development: alterations in lateral root growth (seedlings display 11% reduction in primary root length when grown on normal medium, number of lateral roots is reduced by 25%, length of the lateral roots is reduced by almost 50%); alterations in apical dominance (reduced branching, reduced length of the secondary inflorescences); delayed flowering; reduced plant height; eventually plants grow and produce seeds normally. Enhanced growth under Mn2+ and Mg2+ stress conditions; perturbed hormone sensitivities (roots are resistant to auxin but are tolerant to 1-aminocycloproprane-1-carboxylic acid (an ethylene precursor), abscisic acid, and benzyladenine; sensitive to gibberellic acid and epibrassinolide); modestly sensitive to exogenous Ca2+; exhibit a 50% reduction in tonoplast Ca2+/H+ antiport activity, a 40% reduction in tonoplast V-type H+-translocating ATPase activity, a 36% increase in tonoplast Ca2+-ATPase activity; high capacity transporter responsible for maintaining low cytosolic-free Ca2+ concentrations in plant cells by catalyzing pH gradient-energized vacuolar Ca2+ accumulation. Phenotype curated by ABRC.

 

 

AT2G34780

Description : Encodes a novel protein of unknown function that is essential for embryonic development. Severe loss of function alleles are embryo lethal. Analysis of a partial loss of function allele indicates a role for EMB1611 in regulation of endoreduplication and maintenance of meristem cell fate. It appears to be required for maintaining the CLV-WUS regulatory pathway.

GO Biological Process : cell division, chromatin silencing, cotyledon development, covalent chromatin modification, determination of bilateral symmetry, embryo development ending in seed dormancy, embryo sac egg cell differentiation, embryonic pattern specification, gene silencing by RNA, leaf development, lipid storage, mRNA export from nucleus, maintenance of meristem identity, meristem initiation, meristem maintenance, meristem structural organization, mitotic recombination, negative regulation of DNA endoreduplication, photomorphogenesis, primary shoot apical meristem specification, protein ubiquitination, regulation of cell cycle process, regulation of cell differentiation, regulation of flower development, response to freezing, seed dormancy process, seed germination, seed maturation, sister chromatid cohesion, sugar mediated signaling pathway, vegetative to reproductive phase transition of meristem

 

Задание 9. Вы остановились на 2-х ОРС. Как доказать, что одна из них идентична исследуемому гену? Перечислите необходимые этапы работы.

 

1. Отсеквенировать и сравнить на наличие значимых мутаций.
2. Заказать семена в банке с мутациями в генах-кандидатах, провести тест на аллелизм или тест на комплементарность (клонировать ген дикого типа в мутант)

 

CAPs маркеры

No mispriming library specified

Using 1-based sequence positions

OLIGO start len tm gc% any 3' seq

LEFT PRIMER 121 20 60.08 50.00 4.00 1.00 GCCGGATGATGAAAGTGAGT

RIGHT PRIMER 1022 20 60.41 50.00 6.00 2.00 aagcaaacaggcatgctagg

SEQUENCE SIZE: 1076

INCLUDED REGION SIZE: 1076

 

PRODUCT SIZE: 902, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 0.00

 

1 gtctgtgtgtgtgtatgctttgttttggtgtttgtgatatcattactaagcccacaggtg

 

 

61 acattttggttttgttgttgtgtagGATTTTGATCCGGTAAGGCATATTTTGGAGAATGT

 

 

121 GCCGGATGATGAAAGTGAGTTAGCTTATTTTGAGAAGCAGgtactaacttttaacttctc

>>>>>>>>>>>>>>>>>>>>

 

181 tcaaggaattgaatataaactgatgcgtctattgtgaaaggttaagaatgttgatgaacg

 

 

241 ttttagttagtgagtttaataggttttgtttactgtgtagGCTACCTTGAGGTTGGTACA

 

 

301 GCTGGATAAAGTGGCAGAAACTCTGTCTCACCATGTTATGGAGCATCATGAAGTAATGGg

 

 

361 taagtattcttgaattgagaagatcatatgcgtttggttttgtgagtattaatatttgct

 

 

421 tttggccagTGAAGGGGATGAATTTGGTAAGAGAATTGGAGAAAGATTTGAAGATTGCAA

 

 

481 ATGTCATCTGCAAGgtatgagtttcatattcttaacctaaattttacataaacaatacag

 

 

541 tggttttattttctattttatctattactggtttgacttggttttggctgtttttgtgtg

 

 

601 tttattgccagAATGGAAGACGAAATTTGACTTCTTCTATGAATGAGGCTTCAAGAGATC

 

 

661 TTATTGTACACACTCATTCCAAAAAGAAACAAGCTCTTCTGgtaggcttctcaatttatt

 

 

721 tcagacatctaggctccactgattgtttctttaatctgctttcttgaaatatttcatgtg

 

 

781 ttatagtttccttctgaaatcaccatcaatgtgctctagaactttgtgtacaaaattact

 

 

841 ctgtattatcatctatcagaaagtggccgagtatcttttttgttttaatctctaaagtta

 

 

901 cgaacaggatgtttctagcaagtagtttcttagatacaatctaagtgtaacctagcctaa

 

 

961 ttttttgttttaattgctaaacatactgttgagattattattcctagcatgcctgtttgc

<<<<<<<<<<<<<

 

1021 ttctctggataccatgtaataagagcgacatgaaagtttaatctttttacttgcag

<

 

KEYS (in order of precedence):

>>>>>> left primer

<<<<<< right primer

 

Statistics

con too in in no tm tm high high high

sid many tar excl bad GC too too any 3' poly end

ered Ns get reg GC% clamp low high compl compl X stab ok

Left 2163 0 0 0 1 0 1233 322 0 0 0 19 588

Right 2008 0 0 0 141 0 1360 99 0 3 38 6 361

Pair Stats:

considered 5, ok 5

primer3 release 1.1.4

 

Fragment Sizes for Cut with XmnI
recognition sequence : GAAnnnnTTC

Sorted Fragment Size (bp)

 

Fragment Sizes for Cut with BglII

recognition sequence : AGATCT

Sorted Fragment Size (bp)