SEQUENCE OF ORANGUTAN (PONGO PYGMAEUS) FOXP2 GENE.

Monika Świszczorowska¹, Arleta Lebioda¹, Barbara Świątek², Grażyna Dmochowska¹, Barbara Kosowska³, Tadeusz Dobosz^1
1 Molecular Techniques Laboratory, Wrocław Medical University, Poland
² Institute of Forensic Medicine, Wrocław Medical University, Poland
³ Department of Genetics and Animal Breeding, Wrocław University of Environmental and Life Sciences, Poland

ABSTRACT

Our article concerns on resequencing foxp2 gene, called “speech gene”. It has been already sequenced in human, 7 species of primates, several species of birds, mammals and reptiles. Mutations in human foxp2 locus causes changes in developing of brain parts responsible for speech process. While comparing sequencing results we found some divergences in the orangutans (Pongo pygmaeus) foxp2 sequences. Three research teams published different versions of foxp2 sequence. The effects of this article are results of resequencing foxp2 gene in orangutan and its declaration to GenBank with the access number: DQ789573.

Key words: orangutan, foxp2 gene.

INTRODUCTION

In the GenBank, at the address ncbi.nlm.nih.gov [5], are collected sequencing results of many different genes from many organisms. One of the hundreds of thousands sequencing results collected in this database concerns on the foxp2 gene. In 2001, Lai et al. [2] described mutations of that gene, being effective with changes of development some brain regions responsible for speech process. This discovery was proclaim as an exploration of “gene of speech” what was a large exaggeration. Nevertheless it caused a large interest of laryngologists from many research centers because it appeared as a hope to explain some inexplicable causes of muteness and as a potential possibility to diagnose and predict this disorder. During analysis of the foxp2 mutations in children with delayed speech development [3] we also checked the status of these analysis in other organisms. On the beginning of April 2006 foxp2 gene was already sequence in 7 species of birds, alligator, danio fish, pygmy hippopotamus, bottlenose dolphin, mouse, and 7 primate species (Macaca mulata, Pongo pygmaeus, Gorilla gorilla, Hylobates lar, Pan paniscus, Pan troglotydes and Homo sapiens) [5]. While comparing these sequences we found some divergences among the sequencing results of foxp2 gene in orangutan (Pongo pygmaeus). Three research groups published three differing in details versions of the orangutan foxp2 sequence. It induced us to resequence orangutan foxp2 gene ourselves to explain those divergences.

MATERIAL AND METHODS

Orangutan’s DNA was bought in ECACC company (catalogue No. 019A). Primers for all 16 exons were designed by ourselves and their sequences are as following:

E1F 5’TAA TGG AAG TGT GAA GGT GTA ACG 3’

E1R 5’GCA CTG AGG AAA ACA AAA CTT ACC 3’

E2F 5’TGG GAG TTC TTG TAC ATT GAA GCC 3’

E2R 5’AAG GAC AGG AGT GCT TTA CTA ACC 3’

E3F 5’AAC AGA TAT TTG GTT ATG ACC ACG 3’

E3R 5’GCT AAT AGG TTG TCC TTT CAA ATG 3’

E4F 5’TAT GCC AGT CTA GAA GAG TTT AGG 3’

E4R 5’TGT TAT GTA TCA ATG AGA TAA CCG 3’

E5F 5’TTC TCT GCT GTT TAC TGG TTT GGG 3’

E5R 5’TAA GGC AGA AAG GCC ATG AAA TGG 3’

E6F 5’GTG AAG CTT TCT TTC ATT TTA TAG 3’

E6R 5’TTC TTT TTC GTT GTC TCA ATT GTG 3’

E7F 5’TGA CGT CGT GTT CTT TTG CTA CAG 3’

E7R 5’GCC TAA AAT GCC CAT ATA ATC CAG 3’

E8F 5’GTT TTG TGT CTT CTG TTT GTT TAG 3’

E8R 5’AAG AGT CCA TTA AGT GAG CAT TTA 3’

E9F 5’TCT GAT CTC ACT CTT TCT TAA CAG 3’

E9R 5’CTT TAT AAA GCA ATA TGC ACT TAC 3’

E10F 5’TCC CCT TCT CTT GCG CCT TTG CAG 3’

E10R 5’CTC TAC TGA GCA TTC ATA TGC TAC 3’

E11+12F 5’AAC ACC TAG CTT TTA TTT TTA TAG 3’

E11+12R 5’TAA AAC TGC TGG AAA AGT AGT TAC 3’

E13F 5’TTT CTT CTC TTC TGT CTG CTT TAG 3’

E13R 5’ATT ACA AAA CTA TCA CAA ACA TAC 3’

E14F 5’AGT GTG AGA CAA GCC AGA ACA TAC 3’

E14R 5’GTG TCC TTC TTG CTT CTT CAC ATC 3’

E15F 5’TTT ACA CAA TCT TCA TTT CAC TAG 3’

E15R 5’AGA GAG TCT GTT AAC CAC ACT TAC 3’

E16F 5’TTG GTG TAT CTA CAT GTT TTT CAG 3’

E16R 5’TCC ATG CTT AAT AAA TAG TCA CAG 3’

PCR reactions were performed with Qiagen Multiplex PCR Kit, strict due to the rules of manual. Sequencing PCR reactions were performed with F primers for each exon and for second thread of DNA with R primers to control. Sequencing reactions were performed with genetic analyzer ABI Prism 310 with Big DyeTerminator v 1.1 Cycle Sequencing Kit. Results analysis was executed with computer software ABI Prism Sequencing Analysis v. 3.7.

RESULTS AND DISCUSSION

We compared results of our foxp2 gene sequencing in orangutan to other author results. The differences between these results are shown in tables I to III. Exon 1 sequence is: atg¹ atg² cag³ gaa⁴ tct⁵ gtg⁶ aca⁷ gag⁸ aca⁹ ata¹⁰ agc¹¹ aac¹² agt¹³ tca¹⁴ atg¹⁵ aat¹⁶ caa¹⁷ aat¹⁸ gga¹⁹ atg²⁰ agc²¹ act²² cta²³ agc²⁴ agc²⁵ caa²⁶ tta²⁷ gat²⁸ gct²⁹ ggc³⁰ agc³¹ aga³² gat³³ gga³⁴ aga³⁵ tca³⁶ agt³⁷ ggt³⁸ gac³⁹ acc⁴⁰ agc⁴¹ tct⁴² gaa⁴³ gta⁴⁴ agc⁴⁵ aca⁴⁶ gta⁴⁷ gaa⁴⁸ ctg⁴⁹ ctg⁵⁰ cat⁵¹ ctg⁵² caa⁵³ caa⁵⁴ cag⁵⁵ cag⁵⁶. The codon number was marked over line. Within the whole exon 1 we did not found alleles.

Exons from 2 to 5 are exactly the same both in our and previously published analysis (AF512949, AY143181). Sequences of those exons are as following:

• exon 2 – gct⁵⁷ ctc⁵⁸ cag⁵⁹ gca⁶⁰ gca⁶¹ aga⁶² caa⁶³ ctt⁶⁴ ctt⁶⁵ tta⁶⁶ cag⁶⁷ cag⁶⁸ caa⁶⁹ aca⁷⁰ agt⁷¹ gga⁷² ttg⁷³ aaa⁷⁴ tct⁷⁵ cct⁷⁶ aag⁷⁷ agc⁷⁸ agt⁷⁹ gat⁸⁰ aaa⁸¹ cag⁸² aga⁸³ cca⁸⁴ ctg⁸⁵ cag⁸⁶,

• exon 3 – gtg⁸⁷ cct⁸⁸ gtg⁸⁹ tca⁹⁰ gtg⁹¹ gcc⁹² atg⁹³ atg⁹⁴ act⁹⁵ ccc⁹⁶ cag⁹⁷ gtg⁹⁸ atc⁹⁹ acc¹⁰⁰ cct¹⁰¹ cag¹⁰² caa¹⁰³ atg¹⁰⁴ cag¹⁰⁵ cag¹⁰⁶ atc¹⁰⁷ ctt¹⁰⁸ cag¹⁰⁹ caa¹¹⁰ caa¹¹¹ gtc¹¹² ctg¹¹³ tct¹¹⁴ cct¹¹⁵ cag¹¹⁶ cag¹¹⁷ ctc¹¹⁸ caa¹¹⁹ gcc¹²⁰ ctt¹²¹ ctc¹²² caa¹²³ caa¹²⁴ cag¹²⁵ cag¹²⁶ gcc¹²⁷ gtc¹²⁸ atg¹²⁹ ctg¹³⁰ cag¹³¹ cag¹³²,

• exon 4 – caa¹³³ caa¹³⁴ cta¹³⁵ caa¹³⁶ gag¹³⁷ ttt¹³⁸ tac¹³⁹ aag¹⁴⁰ aaa¹⁴¹ cag¹⁴² caa¹⁴³ gag¹⁴⁴ cag¹⁴⁵ tta¹⁴⁶ cat¹⁴⁷ ctt¹⁴⁸ cag¹⁴⁹ ctt¹⁵⁰ ttg¹⁵¹ cag¹⁵² cag¹⁵³ cag¹⁵⁴ caa¹⁵⁵ cag¹⁵⁶ cag¹⁵⁷ cag¹⁵⁸ cag¹⁵⁹ caa¹⁶⁰ caa¹⁶¹ cag¹⁶² cag¹⁶³ caa¹⁶⁴ caa¹⁶⁵ cag¹⁶⁶ cag¹⁶⁷ cag¹⁶⁸ cag¹⁶⁹ caa¹⁷⁰ caa¹⁷¹ caa¹⁷² caa¹⁷³ cag¹⁷⁴ cag¹⁷⁵ cag¹⁷⁶ caa¹⁷⁷ cag¹⁷⁸ cag¹⁷⁹ cag¹⁸⁰ cag¹⁸¹ cag¹⁸² caa¹⁸³ cag¹⁸⁴ cag¹⁸⁵ cag¹⁸⁶ cag¹⁸⁷ cag¹⁸⁸ caa¹⁸⁹ cag¹⁹⁰ cat¹⁹¹ cct¹⁹² gga¹⁹³ aag¹⁹⁴ caa¹⁹⁵ gcg¹⁹⁶ aaa¹⁹⁷ gag¹⁹⁸.

During analysis of the exon 5 we explained divergences between sequences AF512949 and AY143181. Beginning of the exon 5 is similar in both: cag¹⁹⁹ cag²⁰⁰ cag²⁰¹ cag²⁰² cag²⁰³ cag²⁰⁴. Further we have found difference showed in table no. 1.

Table 1. Differences between orangutan’s foxp2 sequences [1, 6] within exon 5 after codon 204

Source	Codon number and nucleotide sequence
NCBI AY143181 NCBI AF512949 and our results	cag (codon addition between cag204 and caa205) --- (lack of codon)

Due to this result correct sequence was registered by Enard et al. [1] (NCBI access number AF512949). As show in table 1, Hang et al. [6] (NCBI access number AY143181) add to sequence additional codon CAG, between codons 204 and 205. Further part of all exon 5 sequences is the same in both published sequences (AF512949, AY143181) and our results. That part of exon 5 is as following: caa²⁰⁵ cag²⁰⁶ cag²⁰⁷ ttg²⁰⁸ gca²⁰⁹ gcc²¹⁰ cag²¹¹ cag²¹² ctt²¹³ gtc²¹⁴ ttc²¹⁵ cag²¹⁶ cag²¹⁷ cag²¹⁸ ctt²¹⁹ ctc²²⁰ cag²²¹ atg²²² caa²²³ caa²²⁴ ctc²²⁵ cag²²⁶ cag²²⁷ cag²²⁸ cag²²⁹ cat²³⁰ ctg²³¹ ctc²³² agc²³³ ctt²³⁴ cag²³⁵ cgt²³⁶ cag²³⁷ gga²³⁸ ctc²³⁹ atc²⁴⁰ tcc²⁴¹ att²⁴² cca²⁴³ cct²⁴⁴ ggc²⁴⁵ cag²⁴⁶ gca²⁴⁷ gcc²⁴⁸ ctt²⁴⁹ cct²⁵⁰ gtc²⁵¹ caa²⁵² tcg²⁵³ ctg²⁵⁴ cct²⁵⁵ caa²⁵⁶.

When we were analyzing exon 6, a new (third) published foxp2 sequence has appeared in GenBank – AY064615 by Walther et al. [4]. Unfortunately it did not contain all exon 6 sequence, but only part from codon 277 till the end. Nevertheless it was included to exon 6 and further exons analysis. After resequencing exon 6 we did not found any divergences between our and published data from codon 257 to 304 inclusive. Sequence of this fragment is as following: gct²⁵⁷ ggc²⁵⁸ tta²⁵⁹ agt²⁶⁰ cct²⁶¹ gct²⁶² gag²⁶³ att²⁶⁴ cag²⁶⁵ cag²⁶⁶ tta²⁶⁷ tgg²⁶⁸ aaa²⁶⁹ gaa²⁷⁰ gtg²⁷¹ act²⁷² gga²⁷³ gtt²⁷⁴ cac²⁷⁵ agt²⁷⁶ atg²⁷⁷ gaa²⁷⁸ gac²⁷⁹ aat²⁸⁰ ggc²⁸¹ att²⁸² aaa²⁸³ cat²⁸⁴ gga²⁸⁵ ggg²⁸⁶ cta²⁸⁷ gac²⁸⁸ ctc²⁸⁹ act²⁹⁰ act²⁹¹ aac²⁹² aat²⁹³ tcc²⁹⁴ tcc²⁹⁵ tcg²⁹⁶ act²⁹⁷ acc²⁹⁸ tcc²⁹⁹ tcc³⁰⁰ acc³⁰¹ act³⁰² tcc³⁰³ aaa³⁰⁴.

In codon 305 we found one nucleotide polymorphism a/g. It is shown in table 2.

Table 2. SNP a/g type polymorphism in codon 306 of exon 6 resulting from comparison our results with published data [1,4,6]

Source	Codon number and nucleotide sequence
AY064615, AF512949 and our results AY143181	gca305 gcg305

Sequence from 306 to 313 codon is equal in all analyzed data (published and our results) and is as following: tca³⁰⁶ cca³⁰⁷ cca³⁰⁸ ata³⁰⁹ act³¹⁰ cat³¹¹ cat³¹² tcc³¹³. In position 314 codon 6 we found SNP a/c polymorphism, which is shown in table 3.

Table 3. SNP a/c type polymorphism in codon 314 of exon 6 resulting from comparison our results with published data [1,4,6]

Source	Codon number and nucleotide sequence
AY064615, AF512949 and our results AY143181	ata314 atc314

From codon 315 till the end of this exon nucleotide sequence is similar in all analyzed data and is as following: gtg³¹⁵ aat³¹⁶ gga³¹⁷ cag³¹⁸ tct³¹⁹ tca³²⁰ gtt³²¹ cta³²² aat³²³ gca³²⁴ aga³²⁵ cga³²⁶ gac³²⁷ agc³²⁸. Codon 328 was teared by intron and its last nucleotide “C” in reality begins exon 7.

Exon 7 also did not indicate any diversity between our results and those from GenBank. Sequence of exon 7 is as following: tcg³²⁹ tca³³⁰ cat³³¹ gag³³² gag³³³ act³³⁴ ggg³³⁵ gcc³³⁶ tct³³⁷ cac³³⁸ act³³⁹ ctc³⁴⁰ tat³⁴¹ ggc³⁴² cat³⁴³ gga³⁴⁴ gtt³⁴⁵ tgc³⁴⁶ aaa³⁴⁷ tgg³⁴⁸ cca³⁴⁹ ggc³⁵⁰ tgt³⁵¹ gaa³⁵² agc³⁵³ att³⁵⁴ tgt³⁵⁵ gaa³⁵⁶ gat³⁵⁷ ttt³⁵⁸ gga³⁵⁹ cag³⁶⁰ ttt³⁶¹ tta³⁶² aag³⁶³. Last nucleotide of tore codon 363 – “g”, came across in the beginning of exon 8.

Exons from 8 to 19 were exactly the same in all published results and our research, and its sequence was as following:

• exon 8 – cac³⁶⁴ ctt³⁶⁵ aac³⁶⁶ aat³⁶⁷ gaa³⁶⁸ cac³⁶⁹ gca³⁷⁰ ttg³⁷¹ gat³⁷² gac³⁷³ cga³⁷⁴ agc³⁷⁵ act³⁷⁶ gct³⁷⁷ cag³⁷⁸ tgt³⁷⁹ cga³⁸⁰ gtg³⁸¹ caa³⁸² atg³⁸³ cag³⁸⁴ gtg³⁸⁵ gtg³⁸⁶ caa³⁸⁷ cag³⁸⁸ tta³⁸⁹ gaa³⁹⁰ ata³⁹¹ cag³⁹²,

• exon 9 – ctt³⁹³ tct³⁹⁴ aaa³⁹⁵ gaa³⁹⁶ cgc³⁹⁷ gaa³⁹⁸ cgt³⁹⁹ ctt⁴⁰⁰ caa⁴⁰¹ gca⁴⁰² atg⁴⁰³ atg⁴⁰⁴ acc⁴⁰⁵ cac⁴⁰⁶ ttg⁴⁰⁷ cac⁴⁰⁸ atg⁴⁰⁹ cga⁴¹⁰ ccc⁴¹¹ tca⁴¹² gag⁴¹³ ccc⁴¹⁴ aaa⁴¹⁵ cca⁴¹⁶ tct⁴¹⁷ ccc⁴¹⁸ aaa⁴¹⁹ cct⁴²⁰,

• exon 10 – cta⁴²¹ aat⁴²² ctg⁴²³ gtg⁴²⁴ tct⁴²⁵ agt⁴²⁶ gtc⁴²⁷ acc⁴²⁸ atg⁴²⁹ tcg⁴³⁰ aag⁴³¹ aat⁴³² atg⁴³³ ttg⁴³⁴ gag⁴³⁵ aca⁴³⁶ tcc⁴³⁷ cca⁴³⁸ cag⁴³⁹ agc⁴⁴⁰ tta⁴⁴¹ cct⁴⁴² caa⁴⁴³ acc⁴⁴⁴ cct⁴⁴⁵ acc⁴⁴⁶ aca⁴⁴⁷ cca⁴⁴⁸ acg⁴⁴⁹ gcc⁴⁵⁰ cca⁴⁵¹ gtc⁴⁵² acc⁴⁵³ ccg⁴⁵⁴ att⁴⁵⁵ acc⁴⁵⁶ cag⁴⁵⁷ gga⁴⁵⁸ ccc⁴⁵⁹ tca⁴⁶⁰ gta⁴⁶¹ atc⁴⁶² acc⁴⁶³ cca⁴⁶⁴ gcc⁴⁶⁵ agt⁴⁶⁶ gtg⁴⁶⁷ ccc⁴⁶⁸ aat⁴⁶⁹ gtg⁴⁷⁰ gga⁴⁷¹ gcc⁴⁷² ata⁴⁷³ cga⁴⁷⁴ agg⁴⁷⁵ cga⁴⁷⁶ cat⁴⁷⁷ tca⁴⁷⁸ gac⁴⁷⁹ aaa⁴⁸⁰ tac⁴⁸¹ aac⁴⁸² att⁴⁸³ ccc⁴⁸⁴ atg⁴⁸⁵ tca⁴⁸⁶ tca⁴⁸⁷.

Similarly to exons 6 and 7, first codon of exon 11 (codon nr. 488) was amused and its first nucleotide “g” was added on the begging of exon 10. Sequences of exon 11 did not differ between all analyzed data and were as following: gaa⁴⁸⁸ att⁴⁸⁹ gcc⁴⁹⁰ cca⁴⁹¹ aac⁴⁹² tac⁴⁹³ gaa⁴⁹⁴ ttt⁴⁹⁵ tat⁴⁹⁶ aaa⁴⁹⁷ aat⁴⁹⁸ gca⁴⁹⁹ gat⁵⁰⁰ gtc⁵⁰¹ aga⁵⁰² cct⁵⁰³ cca⁵⁰⁴ ttt⁵⁰⁵ act⁵⁰⁶ tat⁵⁰⁷ gca⁵⁰⁸ act⁵⁰⁹ ctc⁵¹⁰ ata⁵¹¹ agg⁵¹² cag⁵¹³.

Exons 12, 13 and 14 had exactly the same sequence both in published data and our research. Sequence of these exons was as following:

• exon 12 – gct⁵¹⁴ atc⁵¹⁵ atg⁵¹⁶ gag⁵¹⁷ tca⁵¹⁸ tct⁵¹⁹ gac⁵²⁰ agg⁵²¹ cag⁵²² tta⁵²³ aca⁵²⁴ ctt⁵²⁵ aat⁵²⁶ gaa⁵²⁷ att⁵²⁸ tac⁵²⁹ agc⁵³⁰ tgg⁵³¹ ttt⁵³² aca⁵³³ cgg⁵³⁴ acg⁵³⁵ ttt⁵³⁶ gct⁵³⁷ tac⁵³⁸ ttc⁵³⁹ agg⁵⁴⁰ cgt⁵⁴¹ aat⁵⁴² gca⁵⁴³ gca⁵⁴⁴ act⁵⁴⁵ tgg⁵⁴⁶ aag⁵⁴⁷,

• exon 13 – aat⁵⁴⁸ gca⁵⁴⁹ gta⁵⁵⁰ cgt⁵⁵¹ cat⁵⁵² aat⁵⁵³ ctt⁵⁵⁴ agc⁵⁵⁵ ctg⁵⁵⁶ cac⁵⁵⁷ aag⁵⁵⁸ tgt⁵⁵⁹ ttt⁵⁶⁰ gtt⁵⁶¹ cga⁵⁶² gta⁵⁶³ gaa⁵⁶⁴ aat⁵⁶⁵ gtt⁵⁶⁶ aaa⁵⁶⁷ gga⁵⁶⁸ gca⁵⁶⁹ gta⁵⁷⁰ tgg⁵⁷¹ act⁵⁷² gtg⁵⁷³ gat⁵⁷⁴ gaa⁵⁷⁵ gta⁵⁷⁶ gaa⁵⁷⁷ tac⁵⁷⁸ cag⁵⁷⁹ aag⁵⁸⁰ cga⁵⁸¹ agg⁵⁸² tca⁵⁸³ caa⁵⁸⁴ aag⁵⁸⁵ ata⁵⁸⁶ aca⁵⁸⁷ gga⁵⁸⁸,

• exon 14 – agt⁵⁸⁹ cca⁵⁹⁰ acc⁵⁹¹ tta⁵⁹² gta⁵⁹³ aaa⁵⁹⁴ aat⁵⁹⁵ ata⁵⁹⁶ cct⁵⁹⁷ acc⁵⁹⁸ agt⁵⁹⁹ tta⁶⁰⁰ ggc⁶⁰¹ tat⁶⁰² gga⁶⁰³ gca⁶⁰⁴ gct⁶⁰⁵ ctt⁶⁰⁶ aat⁶⁰⁷ gcc⁶⁰⁸ agt⁶⁰⁹ ttg⁶¹⁰ cag⁶¹.

Last codon (588) of exon 13 was amused and its first nucleotide was attached to the beginning of exon 14.

Exon 15 almost among whole sequence (when we skip last codon) was similar when we compared data from published and our sequence. Sequence of this codon is as following: gct⁶¹² gcc⁶¹³ ttg⁶¹⁴ gca⁶¹⁵ gag⁶¹⁶ agc⁶¹⁷ agt⁶¹⁸ tta⁶¹⁹ cct⁶²⁰ ttg⁶²¹ cta⁶²² agt⁶²³ aat⁶²⁴ mct⁶²⁵ gga⁶²⁶ ctg⁶²⁷ atc⁶²⁸ aat⁶²⁹ aat⁶³⁰ gca⁶³¹ tcc⁶³² agt⁶³³ ggc⁶³⁴ cta⁶³⁵ ctg⁶³⁶ cag⁶³⁷ gcc⁶³⁸ gtc⁶³⁹ cac⁶⁴⁰ gaa⁶⁴¹ gac⁶⁴² ctc⁶⁴³ aat⁶⁴⁴ ggt⁶⁴⁵ tct⁶⁴⁶ ctg⁶⁴⁷ gat⁶⁴⁸ cac⁶⁴⁹ att⁶⁵⁰ gac⁶⁵¹ agc⁶⁵² aat⁶⁵³ gga⁶⁵⁴ aac⁶⁵⁵ agt⁶⁵⁶ agt⁶⁵⁷ ccg⁶⁵⁸ ggc⁶⁵⁹ tgc⁶⁶⁰ tcg⁶⁶¹ cct⁶⁶² cag⁶⁶³ ccg⁶⁶⁴ cac⁶⁶⁵ ata⁶⁶⁶. Last codon (666) of exon 15 is tore and “a” nucleotide was attached to exon 16. Form of writing codon 625 was taken from AY064615, and was written as mct (m means a or c, but both in our results and sequence AF512949 instead of m is c).

The last exon 16 did not have any differences between our results and published data [1, 4, 6], and its structure is as following: cat⁶⁶⁷ tca⁶⁶⁸ atc⁶⁶⁹ cat⁶⁷⁰ gtc⁶⁷¹ aag⁶⁷² gaa⁶⁷³ gag⁶⁷⁴ cca⁶⁷⁵ gtg⁶⁷⁶ att⁶⁷⁷ gca⁶⁷⁸ gag⁶⁷⁹ gat⁶⁸⁰ gaa⁶⁸¹ gac⁶⁸² tgc⁶⁸³ cca⁶⁸⁴ atg⁶⁸⁵ tcc⁶⁸⁶ tta⁶⁸⁷ gtg⁶⁸⁸ aca⁶⁸⁹ aca⁶⁹⁰ gct⁶⁹¹ aat⁶⁹² cac⁶⁹³ agt⁶⁹⁴ cca⁶⁹⁵ gaa⁶⁹⁶ tta⁶⁹⁷ gaa⁶⁹⁸ gac⁶⁹⁹ gac⁷⁰⁰ aga⁷⁰¹ gag⁷⁰² att⁷⁰³ gaa⁷⁰⁴ gaa⁷⁰⁵ gag⁷⁰⁶ cct⁷⁰⁷ tta⁷⁰⁸ tct⁷⁰⁹ gaa⁷¹⁰ gat⁷¹¹ ctg⁷¹² gaa⁷¹³.

After exon 16 we found non coding part of DNA. This part of DNA did not belong to mRNA sequence – CDS, and as promoteur and introns should not be published. However cited authors published this part [1, 4 and 6] and we decided to do the same. In AY064615, AF512949, AY143181 and also our resuts it is TGA fragment. Moreover in AY064615 it is further fragment GAACTGACTTGTGAAACCTCAGCGTGAAGGGACATATCACTGACCTTCATAACCACTCCACAACAATGAATATTTGACAAATTTTTACTGTGACTATTTATT, which is similar to our results, and the last part which we did not sequence: AAGCATGGAT.

To sum up appropriate CDS sequence of foxp2 gene in Pongo pygmaeus, taking into account all already published and discovered by us alleles, written in fasta format is as following:

atgatgcaggaatctgtgacagagacaataagcaacagttcaatgaatcaaaatggaatgagcactctaagcagccaattagatgctggcagcagagatggaagatcaagtggtgacaccagctctgaagtaagcacagtagaactgctgcatctgcaacaacagcaggctctccaggcagcaag
acaacttcttttacagcagcaaacaagtggattgaaatctcctaagagcagtgataaacagagaccactgcaggtgcctgtgtcagtggccatgatgactccccaggtgatcacccctcagcaaatgcagcagatccttcagcaacaagtcctgtctcctcagcagctccaagcccttctccaacaacag
caggccgtcatgctgcagcagcaacaactacaagagttttacaagaaacagcaagagcagttacatcttcagcttttgcagcagcagcaacagcagcagcagcaacaacagcagcaacaacagcagcagcagcaacaacaacaacagcagcagcaacagcagcagcagcagcaacagcag
cagcagcagcaacagcatcctggaaagcaagcgaaagagcagcagcagcagcagcagcaacagcagttggcagcccagcagcttgtcttccagcagcagcttctccagatgcaacaactccagcagcagcagcatctgctcagccttcagcgtcagggactcatctccattccacctggccaggcag
cccttcctgtccaatcgctgcctcaagctggcttaagtcctgctgagattcagcagttatggaaagaagtgactggagttcacagtatggaagacaatggcattaaacatggagggctagacctcactactaacaattcctcctcgactacctcctccaccacttccaaagcatcaccaccaataactcatcat
tccatagtgaatggacagtcttcagttctaaatgcaagacgagacagctcgtcacatgaggagactggggcctctcacactctctatggccatggagtttgcaaatggccaggctgtgaaagcatttgtgaagattttggacagtttttaaagcaccttaacaatgaacacgcattggatgaccgaagcactgctcag
tgtcgagtgcaaatgcaggtggtgcaacagttagaaatacagctttctaaagaacgcgaacgtcttcaagcaatgatgacccacttgcacatgcgaccctcagagcccaaaccatctcccaaacctctaaatctggtgtctagtgtcaccatgtcgaagaatatgttggagacatccccacagagct
tacctcaaacccctaccacaccaacggccccagtcaccccgattacccagggaccctcagtaatcaccccagccagtgtgcccaatgtgggagccatacgaaggcgacattcagacaaatacaacattcccatgtcatcagaaattgccccaaactacgaattttataaaaatgcagatgtcagacct
ccatttacttatgcaactctcataaggcaggctatcatggagtcatctgacaggcagttaacacttaatgaaatttacagctggtttacacggacgtttgcttacttcaggcgtaatgcagcaacttggaagaatgcagtacgtcataatcttagcctgcacaagtgttttgttcgagtagaaaatgttaaaggagcagtat
ggactgtggatgaagtagaataccagaagcgaaggtcacaaaagataacaggaagtccaaccttagtaaaaaatatacctaccagtttaggctatggagcagctcttaatgccagtttgcaggctgccttggcagagagcagtttacctttgctaagtaatmctggactgatcaataatgcatccagtg
gcctactgcaggccgtccacgaagacctcaatggttctctggatcacattgacagcaatggaaacagtagtccgggctgctcgcctcagccgcacatacattcaatccatgtcaaggaagagccagtgattgcagaggatgaagactgcccaatgtccttagtgacaacagctaatcacagtccagaattag
aagacgacagagagattgaagaagagcctttatctgaagatctggaa

– and in his form was submitted to GenBank NCBI [5], where it received reference number: DQ789573.

REFERENCES

1. Enard W., Przeworski M., Fisher S.E., Lai C.S.L., Wiebe V., Kitano T., Monaco A.P., Pääbo S., 2002. Molecular evolution of FOXP2, a gene involved in speech and language. Nature 418: 869-872 and direct submission to NCBI (AF512949).

2. Lai C.S.L., Fisher S.E., Hurst J.A., Varga-Khadem F., Monaco A.P., 2001. A fokhead-domain gene in mutated ina Severe speech and language disorder. Nature 413:519-523.

3. Mielnik J., 2006. Foxp2 u dzieci z Prostym Opóznionym Rozwojem mowy. Praca doktorska, Akademia Medyczna we Wrocławiu.

4. Walter N.A.R., Thompson J., McGoldrick D., Messier W., 2001. Direct submission to NCBI (AY064615).

5. www.ncbi.nlm.nih.hov

6. Zhang J., Webb D.M., Podlaha O., 2002. Direct submission to NCBI (AY143181).

 

Accepted for print: 7.12.2006

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Monika Świszczorowska
Molecular Techniques Laboratory,
Wrocław Medical University, Poland
M.Curie -Skłodowskiej 52, Wrocław, Poland

Arleta Lebioda
Molecular Techniques Laboratory,
Wrocław Medical University, Poland
M.Curie -Skłodowskiej 52, Wrocław, Poland

Barbara Świątek
Institute of Forensic Medicine,
Wrocław Medical University, Poland
Mikulicza-Radeckiego 4, 50-368 Wrocław, Poland

Grażyna Dmochowska
Molecular Techniques Laboratory,
Wrocław Medical University, Poland
M.Curie -Skłodowskiej 52, Wrocław, Poland

Barbara Kosowska
Department of Genetics and Animal Breeding,
Wrocław University of Environmental and Life Sciences, Poland
Kożuchowska 7, 51-631 Wrocław, Poland
email: basia@gen.ar.wroc.pl

Tadeusz Dobosz
Molecular Techniques Laboratory,
Wrocław Medical University, Poland
M.Curie -Skłodowskiej 52, Wrocław, Poland

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Responses to this article, comments are invited and should be submitted within three months of the publication of the article. If accepted for publication, they will be published in the chapter headed 'Discussions' and hyperlinked to the article.

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