PROJECTUBE3A GENE

• Presented by: Hina Amir

• Official name “ubiquitin protein ligase E3A.”

• Ubiquitin protein ligases target other proteins to be broken down (degraded) within cells.

• Removes damaged or unnecessary proteins.

• Critical role in the normal development and function of the nervous system

Gene Introduction

• Mutations in the UBE3A gene.

• Developmental delays

• Lack of or minimal speech

• Inability to walk

• Trembling movement of arms and legs

• Frequent smiling and laughter

• Happy, excitable personality

• (Happy puppets)

Angelman Syndrome

• Location:15q11.2

• Exons 13

Gene Location

Gene Sequencehttp://www.ncbi.nlm.nih.gov/nuccore/NC_000015.10?report=fa

sta&from=25337245&to=25439043&strand=true

Protein Sequence

• http://www.ncbi.nlm.nih.gov/protein/14424503?report=fasta

http://asia.ensembl.org/Homo_sapiens/Transcript/Sequence_cDNA?db=core;g=ENSG00000114062;r=15:25582381-25684128;t=ENST00000232165

No. of Transcripts

http://asia.ensembl.org/Homo_sapiens/Transcript/Summary?db=core;g=ENSG00000114062;r=15:25582381-25684128;t=ENST00000232165

6 transcripts

5’ and 3’ UTR

• http://asia.ensembl.org/Homo_sapiens/Transcript/Exons?db=core;g=ENSG00000114062;r=15:25582381-25684128;t=ENST00000232165

5’ UTR Repeats

• http://www.repeatmasker.org/tmp/d693122da91a30c1e327e39f8172c2b7.html

3’UTR Repeats

http://www.repeatmasker.org/tmp/ea4c61183827c7237346323b0e36bce6.html

Repeats

http://www.repeatmasker.org/tmp/ea4c61183827c7237346323b0e36bce6.html

SNPs

• There are 1938 active SNPs in human gene UBE3A

http://www.ncbi.nlm.nih.gov/snp/?term=UBE3A

• http://www.ncbi.nlm.nih.gov/projects/SNP/snp_ref.cgi?rs=111033596

Expression

• http://biogps.org/#goto=genereport&id=7337

Multiple Sequence Alignment

• http://www.genome.jp/tools-bin/clustalw

Phylogenetic Tree

• http://www.genome.jp/tools-bin/clustalwtree?tree_upgma+140521141839eYiuH

Primers

LEFT PRIMER

TGGTGACCAATGAATCTCCC

RIGHT PRIMER

GTATAGCCACCGTCATATTCTGT

PRODUCT SIZE: 228

INSILICO PCR

UCSC GENE

http://genome.ucsc.edu/cgi-bin/hgPcr?hgsid=376038365_d1AKZAZ4u4FaQEydv3lKpru0h2o4&org=Human&db=hg38&wp_target=hg38Kgv15&wp_f=TGGTGACCAATGAATCTCCC&wp_r=GTATAGCCACCGTCATATTCTGT&Submit=submit&wp_size=4000&wp_perfect=15&wp_good=15&boolshad.wp_flipReverse=0

Genome Assembly

http://genome.ucsc.edu/cgi-bin/hgPcr?wp_target=&db=hg38&org=Human&wp_f=&wp_r=ATCTAGATTCCGGCTTCCACA&wp_size=4000&wp_perfect=15&wp_good=15&wp_showPage=true&hgsid=376038365_d1AKZAZ4u4FaQEydv3lKpru0h2o4

denaturation at 95°C

Primer annealing at 55°C

Extension at 70°C .

PCR Conditions