PART A
CYP3A4 and CYP3A5 comparative analysis
PCR fragment obtained from CYP3A4 primers. The sequence obtained with the reverse primer is shown below.
CTGTGATCCTAAGAAACCAAATTTTAGGAACTTCTTAGTGCTCTCCACAAAGGGGTCTTGTGGATTGTTGAGAGAGTGAGGTTCACTCCAAATGATGTGCCAGTAATCACATCCATGTTGTTGGCCGACAAAGATGTCTTTCAAGGTGACAGGCTTGCCTTTCTCTGCTTCCCGCCTCAAGTTTCTCACCAATACACCTCCATACTGGGCAATGATGGGGAACATCTCCTTGAGTTTTCCGCTGGGAAGTTGGAAAGCATGCCTTTTTTCATTTTAATCTAACAC
The above sequence was BLAST aligned, at NCBI database.
Human ORFeome Gateway entry vector pENTR223-CYP3A5, complete sequence
Sequence ID: LT741352.1Length: 4296Number of Matches: 1
Related Information
Range 1: 882 to 1139GenBankGraphics Next Match Previous Match
Alignment statistics for match #1Score / Expect / Identities / Gaps / Strand
407 bits(220) / 9e-110 / 248/260(95%) / 7/260(2%) / Plus/Minus
Query 5 GATCCTAAGAAACCAAATTTTAGGAACTTCTTAGTGCTCTCCACAAAGGGGTCTTGTGGA 64
||| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1139 GAT-CTAAGAAACCAAATTTTAGGAACTTCTTAGTGCTCTCCACAAAGGGGTCTTGTGGA 1081
Query 65 TTGTTGAGAGAGT-GAGGTTCACTCCAAATGATGTGCCAGTAATCACATCCATGTTGTTG 123
||||||||||||| || ||||||||||||||||||||||||||||||||||||| ||| |
Sbjct 1080 TTGTTGAGAGAGTCGATGTTCACTCCAAATGATGTGCCAGTAATCACATCCATGCTGTAG 1021
Query 124 GCCGACAAAGATGTCTTTCAAGGTGACAGGCTTGCCTTTCTCTGCTTCCCGCCTCAAGTT 183
||| |||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1020 GCCC-CAAAGATGTCTTTCAAGGTGACAGGCTTGCCTTTCTCTGCTTCCCGCCTCAAGTT 962
Query 184 TCTCACCAATACACCTCCATACTGGGCAATGATGGGGAACATCTCCTTGAGTTTTCCGCT 243
||||||||||||| ||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 961 TCTCACCAATACATCTCCATACTGGGCAATGATGGGGAACATCTCCTTGAGTTTTCCGCT 902
Query 244 GG-GAAG-TTGGA-A-AGCA 259
|| |||| ||||| | ||||
Sbjct 901 GGTGAAGGTTGGAGACAGCA 882
PART B
To determine sequence similarity between CYP3A4 and CYP3A5, their mRNA sequences were aligned using CLUSTALW tools (CLUSTAL 2.1 Multiple Nucleotide Sequence Alignments). The primers used in the PCR amplification were highlighted in the CYP3A4 transcript.
Sequence 1: CYP3A4 2792 bp
Sequence 2: CYP3A5 1755 bp
Start of Pairwise alignments
Aligning...
Sequences (1:2) Aligned. Score: 85.1282
Guide tree file created: [clustalw.dnd]
There are 1 groups
Start of Multiple Alignment
Aligning...
Group 1: Sequences: 2 Score:30274
Alignment Score 10426
CLUSTAL-Alignment file created [clustalw.aln]
CLUSTAL 2.1 multiple sequence alignment (Forward and reverse primers are highlighted.)
CYP3A4 ------AATCACTGCTGTGCAGGGCAGGAAAGCTCCATGCACA
CYP3A5 ATAAATCTTTCAGCAGCTTGGCTGAAGACTGCTGTGCAGGGCAGGGAAGCTCCAGGCAAA
* ****************** ******** *** *
CYP3A4 TAGCCCAGCAAAGAGCAACACAGAGCTGAAAGGAAGACTCAGAGGAGAGAGATAAGTAAG
CYP3A5 CAGCCCAGCAAACAGCAGCACTCAGCTAAAAGGAAGACTCACAGAACACAGTTGAAGAAG
*********** **** *** **** ************* ** * * ** * * ***
CYP3A4 GAAAGTAGTGATGGCTCTCATCCCAGACTTGGCCATGGAAACCTGGCTTCTCCTGGCTGT
CYP3A5 GAAAGTGGCGATGGACCTCATCCCAAATTTGGCGGTGGAAACCTGGCTTCTCCTGGCTGT
****** * ***** ********* * ***** *************************
(Forward primer)
CYP3A4 CAGCCTGGTGCTCCTCTATCTATATGGAACCCATTCACATGGACTTTTTAAGAAGCTTGG
CYP3A5 CAGCCTGGTGCTCCTCTATCTATATGGGACCCGTACACATGGACTTTTTAAGAGACTGGG
*************************** **** * ****************** ** **
CYP3A4 AATTCCAGGGCCCACACCTCTGCCTTTTTTGGGAAATATTTTGTCCTACCATAAGGGCTT
CYP3A5 AATTCCAGGGCCCACACCTCTGCCTTTGTTGGGAAATGTTTTGTCCTATCGTCAGGGTCT
*************************** ********* ********** * * **** *
CYP3A4 TTGTATGTTTGACATGGAATGTCATAAAAAGTATGGAAAAGTGTGGGGCTTTTATGATGG
CYP3A5 CTGGAAATTTGACACAGAGTGCTATAAAAAGTATGGAAAAATGTGGGGAACGTATGAAGG
** * ******* ** ** ***************** ******* ***** **
CYP3A4 TCAACAGCCTGTGCTGGCTATCACAGATCCTGACATGATCAAAACAGTGCTAGTGAAAGA
CYP3A5 TCAACTCCCTGTGCTGGCCATCACAGATCCCGACGTGATCAGAACAGTGCTAGTGAAAGA
***** *********** *********** *** ****** ******************
CYP3A4 ATGTTATTCTGTCTTCACAAACCGGAGGCCTTTTGGTCCAGTGGGATTTATGAAAAGTGC
CYP3A5 ATGTTATTCTGTCTTCACAAATCGAAGGTCTTTAGGCCCAGTGGGATTTATGAAAAGTGC
********************* ** *** **** ** ***********************
CYP3A4 CATCTCTATAGCTGAGGATGAAGAATGGAAGAGATTACGATCATTGCTGTCTCCAACCTT
CYP3A5 CATCTCTTTAGCTGAGGATGAAGAATGGAAGAGAATACGGTCATTGCTGTCTCCAACCTT
******* ************************** **** ********************
CYP3A4 CACCAGTGGAAAACTCAAGGAGATGGTCCCTATCATTGCCCAGTATGGAGATGTGTTGGT
CYP3A5 CACCAGCGGAAAACTCAAGGAGATGTTCCCCATCATTGCCCAGTATGGAGATGTATTGGT
****** ****************** **** *********************** *****
CYP3A4 GAGAAATCTGAGGCGGGAAGCAGAGACAGGCAAGCCTGTCACCTTGAAAGACGTCTTTGG
CYP3A5 GAGAAACTTGAGGCGGGAAGCAGAGAAAGGCAAGCCTGTCACCTTGAAAGACATCTTTGG
****** ****************** ************************* *******
CYP3A4 GGCCTACAGCATGGATGTGATCACTAGCACATCATTTGGAGTGAACATCGACTCTCTCAA
CYP3A5 GGCCTACAGCATGGATGTGATTACTGGCACATCATTTGGAGTGAACATCGACTCTCTCAA
********************* *** **********************************
CYP3A4 CAATCCACAAGACCCCTTTGTGGAAAACACCAAGAAGCTTTTAAGATTTGATTTTTTGGA
CYP3A5 CAATCCACAAGACCCCTTTGTGGAGAGCACTAAGAAGTTCCTAAAATTTGGTTTCTTAGA
************************ * *** ****** * *** ***** *** ** **
(reverse primer)
CYP3A4 TCCATTCTTTCTCTCAATAACAGTCTTTCCATTCCTCATCCCAATTCTTGAAGTATTAAA
CYP3A5 TCCATTATTTCTCTCAATAATACTCTTTCCATTCCTTACCCCAGTTTTTGAAGCATTAAA
****** ************* * ************* * **** ** ****** ******
CYP3A4 TATCTGTGTGTTTCCAAGAGAAGTTACAAATTTTTTAAGAAAATCTGTAAAAAGGATGAA
CYP3A5 TGTCTCTCTGTTTCCAAAAGATACCATAAATTTTTTAAGTAAATCTGTAAACAGAATGAA
* *** * ********* *** * ************ *********** ** *****
CYP3A4 AGAAAGTCGCCTCGAAGATACACAAAAGCACCGAGTGGATTTCCTTCAGCTGATGATTGA
CYP3A5 GAAAAGTCGCCTCAACGACAAACAAAAGCACCGACTAGATTTCCTTCAGCTGATGATTGA
*********** * ** * ************* * ***********************
CYP3A4 CTCTCAGAATTCAAAAGAAACTGAGTCCCACAAAGCTCTGTCCGATCTGGAGCTCGTGGC
CYP3A5 CTCCCAGAATTCGAAAGAAACTGAGTCCCACAAAGCTCTGTCTGATCTGGAGCTCGCAGC
*** ******** ***************************** ************* **
CYP3A4 CCAATCAATTATCTTTATTTTTGCTGGCTATGAAACCACGAGCAGTGTTCTCTCCTTCAT
CYP3A5 CCAGTCAATAATCTTCATTTTTGCTGGCTATGAAACCACCAGCAGTGTTCTTTCCTTCAC
*** ***** ***** *********************** *********** *******
CYP3A4 TATGTATGAACTGGCCACTCACCCTGATGTCCAGCAGAAACTGCAGGAGGAAATTGATGC
CYP3A5 TTTATATGAACTGGCCACTCACCCTGATGTCCAGCAGAAACTGCAAAAGGAGATTGATGC
* * ***************************************** **** ********
CYP3A4 AGTTTTACCCAATAAGGCACCACCCACCTATGATACTGTGCTACAGATGGAGTATCTTGA
CYP3A5 AGTTTTGCCCAATAAGGCACCACCTACCTATGATGCCGTGGTACAGATGGAGTACCTTGA
****** ***************** ********* * *** ************* *****
CYP3A4 CATGGTGGTGAATGAAACGCTCAGATTATTCCCAATTGCTATGAGACTTGAGAGGGTCTG
CYP3A5 CATGGTGGTGAATGAAACACTCAGATTATTCCCAGTTGCTATTAGACTTGAGAGGACTTG
****************** *************** ******* ************ **
CYP3A4 CAAAAAAGATGTTGAGATCAATGGGATGTTCATTCCCAAAGGGGTGGTGGTGATGATTCC
CYP3A5 CAAGAAAGATGTTGAAATCAATGGGGTATTCATTCCCAAAGGGTCAATGGTGGTGATTCC
*** *********** ********* * *************** ***** *******
CYP3A4 AAGCTATGCTCTTCACCGTGACCCAAAGTACTGGACAGAGCCTGAGAAGTTCCTCCCTGA
CYP3A5 AACTTATGCTCTTCACCATGACCCAAAGTACTGGACAGAGCCTGAGGAGTTCCGCCCTGA
** ************* **************************** ****** ******
CYP3A4 AAGATTCAGCAAGAAGAACAAGGACAACATAGATCCTTACATATACACACCCTTTGGAAG
CYP3A5 AAGGTTCAGTAAGAAGAA---GGACAGCATAGATCCTTACATATACACACCCTTTGGAAC
*** ***** ******** ***** ********************************
CYP3A4 TGGACCCAGAAACTGCATTGGCATGAGGTTTGCTCTCATGAACATGAAACTTGCTCTAAT
CYP3A5 TGGACCCAGAAACTGCATTGGCATGAGGTTTGCTCTCATGAACATGAAACTTGCTCTAAT
************************************************************
CYP3A4 CAGAGTCCTTCAGAACTTCTCCTTCAAACCTTGTAAAGAAACACAGATCCCCCTGAAATT
CYP3A5 CAGAGTCCTTCAGAACTTCTCCTTCAAACCTTGTAAAGAAACACAGATCCCCTTGAAATT
**************************************************** *******
CYP3A4 AAGCTTAGGAGGACTTCTTCAACCAGAAAAACCCGTTGTTCTAAAGGTTGAGTCAAGGGA
CYP3A5 AGACACGCAAGGACTTCTTCAACCAGAAAAACCCATTGTTCTAAAGGTGGATTCAAGAGA
* * ************************* ************* ** ***** **
CYP3A4 TGGCACCGTAAGTGGAGCCTGAATTTTCCTAAGGACTTCTGCTTTGCTCTTCAAGAAATC
CYP3A5 TGGAACCCTAAGTGGAGAATGAGTTATTCTAAGGATTTCTACTTTGGTCTTCAAGAAAGC
*** *** ********* *** ** * ******* **** ***** *********** *
CYP3A4 TGTGCCTGAGAACACCAGAGACCTCAAATTACTTTGTGAATAGAACTCTGAAATGAAGAT
CYP3A5 TGTGCCCCAGAACACCAGAGATTTCAA----CTTAGTCAATAAAACCTTGAAATAAAGAT
****** ************* **** *** ** **** *** ****** *****
CYP3A4 GGGCTTCATCCAATGGACTGCATAAATAACCGGGGATTCTGTACATGCATTGAGCTCTCT
CYP3A5 GGGCTTAATCTAATGTACTGCA------
****** *** **** ******
CYP3A4 CATTGTCTGTGTAGAGTGTTATACTTGGGAATATAAAGGAGGTGACCAAATCAGTGTGAG
CYP3A5 ------
CYP3A4 GAGGTAGATTTGGCTCCTCTGCTTCTCACGGGACTATTTCCACCACCCCCAGTTAGCACC
CYP3A5 ------
CYP3A4 ATTAACTCCTCCTGAGCTCTGATAAGAGAATCAACATTTCTCAATAATTTCCTCCACAAA
CYP3A5 ------
CYP3A4 TTATTAATGAAAATAAGAATTATTTTGATGGCTCTAACAATGACATTTATATCACATGTT
CYP3A5 ------
CYP3A4 TTCTCTGGAGTATTCTATAAGTTTTATGTTAAATCAATAAAGACCACTTTACAAAAGTAT
CYP3A5 ------
CYP3A4 TATCAGATGCTTTCCTGCACATTAAGGAGAAATCTATAGAACTGAATGAGAACCAACAAG
CYP3A5 ------
CYP3A4 TAAATATTTTTGGTCATTGTAATCACTGTTGGCGTGGGGCCTTTGTCAGAACTAGAATTT
CYP3A5 ------
CYP3A4 GATTATTAACATAGGTGAAAGTTAATCCACTGTGACTTTGCCCATTGTTTAGAAAGAATA
CYP3A5 ------
CYP3A4 TTCATAGTTTAATTATGCCTTTTTTGATCAGGCACAGTGGCTCACGCCTGTAATCCTAGC
CYP3A5 ------
CYP3A4 AGTTTGGGAGGCTGAGCCGGGTGGATCGCCTGAGGTCAGGAGTTCAAGACAAGCCTGGCC
CYP3A5 ------
CYP3A4 TACATGGTTGAAACCCCATCTCTACTAAAAATACACAAATTAGCTAGGCATGGTGGACTC
CYP3A5 ------
CYP3A4 GCCTGTAATCTCACTACACAGGAGGCTGAGGCAGGAGAATCACTTGAACCTGGGAGGCGG
CYP3A5 ------
CYP3A4 ATGTTGAAGTGAGCTGAGATTGCACCACTGCACTCCAGTCTGGGTGAGAGTGAGACTCAG
CYP3A5 ------
CYP3A4 TCTTAAAAAAATATGCCTTTTTGAAGCACGTACATTTTGTAACAAAGAACTGAAGCTCTT
CYP3A5 ------
CYP3A4 ATTATATTATTAGTTTTGATTTAATGTTTTCAGCCCATCTCCTTTCATATTTCTGGGAGA
CYP3A5 ------
CYP3A4 CAGAAAACATGTTTCCCTACACCTCTTGCATTCCATCCTCAACACCCAACTGTCTCGATG
CYP3A5 ------
CYP3A4 CAATGAACACTTAATAAAAAACAGTCGATTGGTCAATTGATTGAGCAATAAGCCT
CYP3A5 ------
To determine the protein similarities between CYP3A4 and CYP4A5, protein sequences were aligned using CLUSTALW tool.
CLUSTAL 2.1 Multiple protein Sequence Alignments
Sequence 1: CYP3A4 503 aa
Sequence 2: CYP3A5 502 aa
Start of Pairwise alignments
Aligning...
Sequences (1:2) Aligned. Score: 83.6653
Guide tree file created: [clustalw.dnd]
There are 1 groups
Start of Multiple Alignment
Aligning...
Group 1: Sequences: 2 Score:7602
Alignment Score 2691
CLUSTAL-Alignment file created [clustalw.aln]
clustalw.aln
CLUSTAL 2.1 multiple sequence alignment
CYP3A4 MALIPDLAMETWLLLAVSLVLLYLYGTHSHGLFKKLGIPGPTPLPFLGNILSYHKGFCMF
CYP3A5 MDLIPNLAVETWLLLAVSLVLLYLYGTRTHGLFKRLGIPGPTPLPLLGNVLSYRQGLWKF
* ***:**:******************::*****:**********:***:***::*: *
CYP3A4 DMECHKKYGKVWGFYDGQQPVLAITDPDMIKTVLVKECYSVFTNRRPFGPVGFMKSAISI
CYP3A5 DTECYKKYGKMWGTYEGQLPVLAITDPDVIRTVLVKECYSVFTNRRSLGPVGFMKSAISL
* **:*****:** *:** *********:*:***************.:***********:
CYP3A4 AEDEEWKRLRSLLSPTFTSGKLKEMVPIIAQYGDVLVRNLRREAETGKPVTLKDVFGAYS
CYP3A5 AEDEEWKRIRSLLSPTFTSGKLKEMFPIIAQYGDVLVRNLRREAEKGKPVTLKDIFGAYS
********:****************.*******************.********:*****
CYP3A4 MDVITSTSFGVNIDSLNNPQDPFVENTKKLLRFDFLDPFFLSITVFPFLIPILEVLNICV
CYP3A5 MDVITGTSFGVNIDSLNNPQDPFVESTKKFLKFGFLDPLFLSIILFPFLTPVFEALNVSL
*****.*******************.***:*:*.****:**** :**** *::*.**:.:
CYP3A4 FPREVTNFLRKSVKRMKESRLEDTQKHRVDFLQLMIDSQNSKETESHKALSDLELVAQSI
CYP3A5 FPKDTINFLSKSVNRMKKSRLNDKQKHRLDFLQLMIDSQNSKETESHKALSDLELAAQSI
**::. *** ***:***:***:*.****:**************************.****
CYP3A4 IFIFAGYETTSSVLSFIMYELATHPDVQQKLQEEIDAVLPNKAPPTYDTVLQMEYLDMVV
CYP3A5 IFIFAGYETTSSVLSFTLYELATHPDVQQKLQKEIDAVLPNKAPPTYDAVVQMEYLDMVV
**************** :**************:***************:*:*********
CYP3A4 NETLRLFPIAMRLERVCKKDVEINGMFIPKGVVVMIPSYALHRDPKYWTEPEKFLPERFS
CYP3A5 NETLRLFPVAIRLERTCKKDVEINGVFIPKGSMVVIPTYALHHDPKYWTEPEEFRPERFS
********:*:****.*********:***** :*:**:****:*********:* *****
CYP3A4 KKNKDNIDPYIYTPFGSGPRNCIGMRFALMNMKLALIRVLQNFSFKPCKETQIPLKLSLG
CYP3A5 KK-KDSIDPYIYTPFGTGPRNCIGMRFALMNMKLALIRVLQNFSFKPCKETQIPLKLDTQ
** **.**********:****************************************.
CYP3A4 GLLQPEKPVVLKVESRDGTVSGA
CYP3A5 GLLQPEKPIVLKVDSRDGTLSGE
********:****:*****:**