Ancestral sequence reconstruction (ASR) – also known as ancestral gene/sequence reconstruction/resurrection – is a technique used in the study of molecular evolution. The method uses related sequences to reconstruct an "ancestral" gene from a multiple sequence alignment.
The method can be used to resurrect ancestral proteins and was suggested in 1963 by Linus Pauling and Emile Zuckerkandl.  In the case of enzymes, this approach has been called paleoenzymology (British: palaeoenzymology). Some early efforts were made in the 1980s and 1990s, led by the laboratory of Steven A. Benner, showing the potential of this technique. Thanks to the improvement of algorithms and of better sequencing and synthesis techniques, the method was developed further in the early 2000s to allow the resurrection of a greater variety of and much more ancient genes. Over the last decade, ancestral protein resurrection has developed as a strategy to reveal the mechanisms and dynamics of protein evolution.