BOBCOTM

Standard DNA Control Sequences


Control Sequences for Sickle Cell Anemia
Genetic Diagnosis Kit

Human beta globin and sickle cell mutant

humhbb.gb_pr is the Human beta globin region on chromosome 11. The sickle cell mutation is a->t at base 62206 of that sequence. The region shown here is a PCR product amplified from the 5' region of the genomic sequence, as described by Husein et al [1]..

Primers:

Regular Expression Notes

Mutation:
TCCTGAGGAG
     ^
A->T, base 62202 of humhbb.gb_pr

Sequences:

> Amplified region of human beta hemoglobin, using the primers of
> Husain et al. Note that the last two bases of this amplified
> product have been mutagenized by the primer.

5-TCCTAAGCCAGTGCCAGAAGAGCCAAGGACAGGTACGGCTGTCATCACTTAGACCTCACCCTGTGGAGCCACACCCTAGGGTTGGCCAATCTACTCCCAGGAGCAGGGAGGGCAGGAGCCAGGGCTGGGCATAAAAGTCAGGGCAGAGCCATCTATTGCTTACATTTGCTTCTGACACAACTGTGTTCACTAGCAACCTCAAACAGACACCATGGTGCACCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAGTTGGTGGTGAGGCCCTGGGCAGGTTGGTATCAAGGTTACAAGACAGGTTTAAGGAGACCAATAGAAACTGGGCATGTGGAGACAGAGAAGACTCTTGGGTTTCTGATAGGCACTGACTCTCTCTGCCTATTGGTCTATTTTCCCACCCTTAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGGGGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGTGCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACTGTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGGTGAGTCTATGGGACCCTTGATGTTTTCTTTCCCCTTCTTTTCTATGGTTAAGTTCATGTCATAGGAAGGGGAGAAGTAACAGGGTACAGTTTAGAATGGGAAACAGACGAATTC-3
> Amplified region of mutant (sickle cell) human beta hemoglobin,
> using the primers of Husain et al. Note that the last two bases
> of this amplified product have been mutagenized by the primer.
5-TCCTAAGCCAGTGCCAGAAGAGCCAAGGACAGGTACGGCTGTCATCACTTAGACCTCACCCTGTGGAGCCACACCCTAGGGTTGGCCAATCTACTCCCAGGAGCAGGGAGGGCAGGAGCCAGGGCTGGGCATAAAAGTCAGGGCAGAGCCATCTATTGCTTACATTTGCTTCTGACACAACTGTGTTCACTAGCAACCTCAAACAGACACCATGGTGCACCTGACTCCTGTGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAGTTGGTGGTGAGGCCCTGGGCAGGTTGGTATCAAGGTTACAAGACAGGTTTAAGGAGACCAATAGAAACTGGGCATGTGGAGACAGAGAAGACTCTTGGGTTTCTGATAGGCACTGACTCTCTCTGCCTATTGGTCTATTTTCCCACCCTTAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGGGGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGTGCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACTGTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGGTGAGTCTATGGGACCCTTGATGTTTTCTTTCCCCTTCTTTTCTATGGTTAAGTTCATGTCATAGGAAGGGGAGAAGTAACAGGGTACAGTTTAGAATGGGAAACAGACGAATTC-3

Reference:


GenBank File for Human Beta-Globin mRNA

LOCUS       HSBGLX        549 bp    mRNA            PRI       17-NOV-1993
DEFINITION  Human messenger RNA for beta-globin.
ACCESSION   V00500
NID         g29445
VERSION     V00500.1  GI:29445
KEYWORDS    beta-globin; complementary DNA; globin.
SOURCE      human.
  ORGANISM  Homo sapiens
            Eukaryota; Metazoa; Chordata; Vertebrata; Mammalia; Eutheria;
            Primates; Catarrhini; Hominidae; Homo.
REFERENCE   1  (bases 1 to 549)
  AUTHORS   Marotta,C.A., Wilson,J.T., Forget,B.G. and Weissman,S.M.
  TITLE     Human beta-globin messenger RNA. III. Nucleotide sequences derived
            from complementary DNA
  JOURNAL   J. Biol. Chem. 252 (14), 5040-5053 (1977)
  MEDLINE   77207124
FEATURES             Location/Qualifiers
     source          1..549
                     /organism="Homo sapiens"
                     /db_xref="taxon:9606"
     mRNA            1..549
                     /note="messenger RNA"
     CDS             1..444
                     /note="reading frame (beta-globin)"
                     /codon_start=1
                     /protein_id="CAA23759.1"
                     /db_xref="PID:g29446"
                     /db_xref="GI:29446"
                     /db_xref="SWISS-PROT:P02023"
                     /translation="MVHLTPVEKSAVTAXWGKVNVDEVGGEALGRLLVVYPWTQRFFE
                     SFGDLSTPDAVMGNPKVKAHGKKVLGAFSDGLAHLDNLKGTFATLSELHCDKLHVDPE
                     NFRLLGNVLVCVLAHHFGKEFTPPVQAAYQKVVAGVANALAHKYH"
BASE COUNT      110 a    135 c    157 g    141 t      6 others
ORIGIN      
        1 atggtgcacc tgactcctgt ggagaagtcy gcngttactg cnytntgggg caaggtgaac
       61 gtggatgaag ttggtggtga ggccctgggc aggctgctgg tggtctaccc ttggacccag
      121 aggttctttg agtcctttgg ggatctgtcc actcctgatg cagttatggg caaccctaag
      181 gtgaaggctc atggcaagaa agtgctcggt gcctttagtg atggcctggc tcacctggac
      241 aacctcaagg gcacctttgc cacactgagt gagctgcact gtgacaagct gcacgtggat
      301 cctgagaact tcaggctcct gggcaacgtg ctggtctgtg tgctggccca tcactttggc
      361 aaagaattca ccccaccagt gcaggcngcc tatcagaaag tggtggctgg tgtggctaat
      421 gccctggccc acaagtatca ctaagctcgc tttcttgctg tccaatttct attaaaggtt
      481 cctttgttcc ctaagtccaa ctactaaact gggggatatt atgaagggcc ttgagcatct
      541 ggattctgc
//

Partial GenBank Human Beta Globin Genomic Sequence

LOCUS       HUMHBB      73308 bp    DNA             PRI       18-FEB-1994
DEFINITION  Human beta globin region on chromosome 11.
ACCESSION   U01317 J00179 J00093 J00094 J00096 J00158 J00159 J00160 J00161
            J00162 J00163 J00164 J00165 J00166 J00167 J00168 J00169 J00170
            J00171 J00172 J00173 J00174 J00175 J00177 J00178 K01239 K01890
            K02544 M18047 M19067 M24868 M24886 X00423 X00424 X00672
NID         g455025
VERSION     U01317.1  GI:455025
KEYWORDS    Alu repeat; HPFH; KpnI repetitive sequence; RNA polymerase III;
            allelic variation; alternate cap site; beta-1 pseudogene;
            beta-globin; delta-globin; epsilon-globin; gamma-globin; gene
            duplication; globin; polymorphism; promoter mutation; pseudogene;
            repetitive sequence; thalassemia.
SOURCE      Human mRNA, cDNA and DNA.
  ORGANISM  Homo sapiens
            Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Mammalia;
            Eutheria; Primates; Catarrhini; Hominidae; Homo.
[...]
     CDS             join(62187..62278,62409..62631,63482..63610)
                     /note="beta-globin"
                     /codon_start=1
                     /protein_id="AAA16334.1"
                     /db_xref="PID:g455997"
                     /db_xref="GI:455997"
                     /translation="MVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFE
                     SFGDLSTPDAVMGNPKVKAHGKKVLGAFSDGLAHLDNLKGTFATLSELHCDKLHVDPE
                     NFRLLGNVLVCVLAHHFGKEFTPPVQAAYQKVVAGVANALAHKYH"
[...]
BASE COUNT    22068 a  14146 c  14785 g  22309 t
ORIGIN      1 bp upstream of EcoRI site; chromosome 11p15 [J. Biol. Chem. 260].
[...]
    62161 ttcactagca acctcaaaca gacaccatgg tgcacctgac tcctgaggag aagtctgccg
    62221 ttactgccct gtggggcaag gtgaacgtgg atgaagttgg tggtgaggcc ctgggcaggt
    62281 tggtatcaag gttacaagac aggtttaagg agaccaatag aaactgggca tgtggagaca
    62341 gagaagactc ttgggtttct gataggcact gactctctct gcctattggt ctattttccc
    62401 acccttaggc tgctggtggt ctacccttgg acccagaggt tctttgagtc ctttggggat
    62461 ctgtccactc ctgatgctgt tatgggcaac cctaaggtga aggctcatgg caagaaagtg
    62521 ctcggtgcct ttagtgatgg cctggctcac ctggacaacc tcaagggcac ctttgccaca
    62581 ctgagtgagc tgcactgtga caagctgcac gtggatcctg agaacttcag ggtgagtcta
    62641 tgggaccctt gatgttttct ttccccttct tttctatggt taagttcatg tcataggaag
    62701 gggagaagta acagggtaca gtttagaatg ggaaacagac gaatgattgc atcagtgtgg
    62761 aagtctcagg atcgttttag tttcttttat ttgctgttca taacaattgt tttcttttgt
    62821 ttaattcttg ctttcttttt ttttcttctc cgcaattttt actattatac ttaatgcctt
    62881 aacattgtgt ataacaaaag gaaatatctc tgagatacat taagtaactt aaaaaaaaac
    62941 tttacacagt ctgcctagta cattactatt tggaatatat gtgtgcttat ttgcatattc
    63001 ataatctccc tactttattt tcttttattt ttaattgata cataatcatt atacatattt
    63061 atgggttaaa gtgtaatgtt ttaatatgtg tacacatatt gaccaaatca gggtaatttt
    63121 gcatttgtaa ttttaaaaaa tgctttcttc ttttaatata cttttttgtt tatcttattt
    63181 ctaatacttt ccctaatctc tttctttcag ggcaataatg atacaatgta tcatgcctct
    63241 ttgcaccatt ctaaagaata acagtgataa tttctgggtt aaggcaatag caatatttct
    63301 gcatataaat atttctgcat ataaattgta actgatgtaa gaggtttcat attgctaata
    63361 gcagctacaa tccagctacc attctgcttt tattttatgg ttgggataag gctggattat
    63421 tctgagtcca agctaggccc ttttgctaat catgttcata cctcttatct tcctcccaca
    63481 gctcctgggc aacgtgctgg tctgtgtgct ggcccatcac tttggcaaag aattcacccc
    63541 accagtgcag gctgcctatc agaaagtggt ggctggtgtg gctaatgccc tggcccacaa
    63601 gtatcactaa gctcgctttc ttgctgtcca atttctatta aaggttcctt tgttccctaa
[...]
Full GenBank Entry